Brochure - BD Biosciences Neuroscience Research Source
Transcript of Brochure - BD Biosciences Neuroscience Research Source
BD Biosciences Neuroscience Research SourceBright Ideas for Neuroscience Research
Abbreviations
Reactivity:
Bab: Baboon; Bov: Bovine; C: Chicken; Cyno: Cynomolgus; D: Dog; Dros: Drosophila; F: Frog; G Pig: Guinea Pig; Gt: Goat; Ham: Hamster; Hs: Horse; Hu: Human; Koa: Koala; Mam: Mammalian; Ms: Mouse; Pig: Pigeon; Qua: Quail; R: Rat; Rb: Rabbit; Rhe: Rhesus; Tr: Trout; VTC: Viral transformed cells; Xen: Xenopus
Applications:
Block: Blocking; FA: Functional Assay; FCM: Flow Cytometry; IC/FCM: Intracellular Flow Cytometry; IF: Immunofluorescence; IHC: Immunohistochemistry; IHC(Fr): Frozen; IHC(F): Formalin; IHC(P): Paraffin; IHC(Zn): Zinc; IP: Immunoprecipitation; Neu: Neutralizing; RIA: Radioimmunoassay; RPA: Ribonuclease Protection Assay; WB: Western blot
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Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5Review Article on the Field of Neuroscience . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Advanced Culture Surfaces for Neuronal Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
BD Falcon Cultureware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
BD Falcon Pipets and Tubes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
BD Primaria Cultureware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Extracellular Matrix Proteins and Attachment Factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
BD Matrigel Basement Membrane Matrix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
BD BioCoat Cellware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
BD BioCoat Matrigel™ Matrix Cellware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
BD BioCoat Laminin Cellware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
BD BioCoat Collagen IV Cellware. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
BD BioCoat Poly-Lysine Cellware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
BD BioCoat Poly-D-Lysine/Laminin, Poly-L-Orinthine/Laminin, and Laminin/Fibronectin Cellware . . . . . . . . . . . . . . . . . 25
BD BioCoat Collagen I Cellware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
BD BioCoat Fibronectin Cellware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
BD BioCoat Gelatin Cellware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
Cell Culture Inserts and Insert Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
BD BioCoat Services . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
BD Growth Factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
Functional Assays and Discovery Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32Apoptosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
Cell Signaling, including Kinases and Phosphotases. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
Spotlight Phosphoprotein Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
BD Phosflow Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
Cytoskeletal Proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
Development and Differentiation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Growth Factor Related . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Ion Channels and Transporters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
Neurotransmitter Receptor-Related . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
Synaptic Vesicles and Post-Synaptic Proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
BD Assay Kits and Reagents for Bioimaging and Flow Cytometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
BD Cytometric Bead Array Flex Set Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
Technical Spotlight and Protocols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60Screening and Development of Antibodies for use in Cell-Based Assays using Immunofluorescence Microscopy . . . . . . . . . 60
Phospho-Protein Profiling in Stem Cells. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
IHC for Neuroscience Research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
Phospho-Specific Staining of Rat Brain Tissues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
Imaging and High Content Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66Bioimaging Instrumentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
Cellular Analysis Using Flow Cytometry. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
BD Flow Cytometry Instrumentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
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Table of Contents
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BD Biosciences Neuroscience Source
• Dependable innovation in cell culture technology for over 50 years
• More than 20 years of cell separation leadership
• Advanced functional assays
• Thousands of reagents and multiple systems for cell analysis
BD Biosciences provides integrated,
high-value products and services for cell
culture and analysis with hallmark brands
such as BD FACS™ instruments, which are
considered the gold standard in the
field of flow cytometry. Along with
BD Pharmingen™ and BD Transduction
Laboratories™ brand antibodies, we offer
one of the largest collections of reagents for
flow cytometric analysis, western blotting,
immunofluorescence microscopy, and ELISA.
With BD Falcon™, the industry’s premier
cultureware family, and the addition
of new instrument platforms such
as BD Pathway™ bioimager and
BD CARV II™ confocal imager, we now
provide one of the most comprehensive
portfolios of cell culture and analysis tools
for the research community. With this
portfolio, BD Biosciences offers tools for
researchers investigating the mechanisms
of neuronal differentiation, growth, and
disease states.
Recognized for its outstanding quality,
consistency, and value for more than 50
years, BD Falcon products have enabled
scientists worldwide to advance their cell
biology research. Building on that
foundation is BD BioCoat™ cellware,
the first commercial pre-coated cell
cultureware combining plastics with
extracellular matrix proteins and attachment
factors. This marks a new era in cell culture
research, making culture and analysis more
efficient and convenient for researchers.
BD Biosciences continues to develop
an integrated approach to neurobiology
research through its reagents, cultureware,
and cellular analysis instrumentation.
This combined approach yields
unprecedented information on neuronal cell
performance, from cell culturing to
high-content analysis. BD Biosciences is
helping to elucidate the inner world of the
cell with new antibody specificities, dyes,
instrumentation, software, and
culturing systems.
Visit bdbiosciences .com/neuroscience
for more information on the latest
neuroscience research tools, product
releases, and technical updates.
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Introduction
Neuroscience is a rapidly changing and expanding area of research with the goal of
deciphering the brain’s neurological systems and related disorders, such as
Parkinson’s and Alzheimer’s diseases. BD Biosciences is proud to be an integral participant
in this important research.
Imagingand
High ContentAnalysis
Cell Culture
Functional Assaysand Discovery Reagents
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Review Article on the Field of Neuroscience
Benjamin Wolozin, MD, PhDProfessor of PharmacologyDepartment of Pharmacology and Experimental TherapeuticsBoston University School of MedicineBoston, MA, USA
A decade of research has led to dramatic
advances in our understanding of the
causes of many diseases of the central
nervous system. One of the striking
observations is the presence of common
biochemical pathways that contribute to
the pathophysiology of a wide variety of
diseases. Neurons, glia and oligodendro-
cytes subjected to varying types of stress
or injury show activation of many common
pathways such as the stress kinase cascades
and the apoptotic cascades. Identification
of these common signaling pathways
provides researchers with the biochemical
framework to explore disease pathways
for known illnesses and new genetic vari-
ants of illnesses. In some cases the central
pathways of disease are known, but the
small tributaries that subtly differentiate
diseases, particularly late onset diseases,
are not understood. Many of these
signaling processes often occur in parallel.
At the same time, fundamental questions
remain. The subtle reasons why one person
develops Alzheimer’s disease, a second
person develops Parkinson’s disease, and
a third is still healthy at 95 years of age
remains a mystery.
Proteins causing familial neurodegenerative diseases
Common diseases of the central nervous
system fall into three general categories:
late onset neurodegenerative diseases, acute
injury, and psychiatric illnesses. The response
of the brain to neurodegenerative diseases
such as Alzheimer’s disease, Parkinson’s
disease, Huntington’s disease and amyo-
trophic lateral sclerosis is pleitropic. Familial
forms of each of these diseases are associ-
ated with abnormalities in a specific protein.
Alzheimer’s disease: Alzheimer’s disease
(AD) is characterized by the accumulation
of neuritic plaques containing aggregated
b-amyloid (Ab) and neurofibrillary tangles
containing fibrillar microtubule associated
protein tau. In AD, mutations in genes that
regulate production of Ab lead to early
onset familial AD. Mutations in amyloid
precursor protein, presenilin 1 and presenilin
2 all cause familial AD. Discovery of these
genes also lead to discovery of the b and
g-secretase complexes. b-Secretase activity
is largely attributed to the action of one
membrane associated protein, BACE 1.
g-Secretase is composed of at least four
different membrane associated proteins:
presenilin, nicastrin, pen-2 and aph-1.
Increased production of total Ab or Ab42,
a longer form of Ab that aggregates
rapidly, leads to increased deposition and
causes neurotoxicity. This neurotoxicity is
accompanied by tau hyper-phosphorylation
and formation of neurofibrillary tangles
composed of tau protein. Proper functioning
of tau is clearly important for neuronal
health because mutations in tau are
associated with a different type of dementia,
termed fronto-temporal dementia.
One of the challenges facing researchers
investigating AD is the distinction between
the uncommon forms of early onset, familial
AD that are associated with mutations in
presenilin or APP, and the more common
forms of disease that tend to occur later
in life. The apolipoprotein E4 isoform is
the only polymorphism clearly associated
with late onset AD. Polymorphisms in other
proteins, such as the ubiquilin 1 protein,
might be associated with late onset AD, but
the associations remain controversial and
the strength of the effect is far below that
of apolipoprotein E4.
Parkinson’s disease: At least 5 different
genes have been associated with familial
Parkinson’s disease (PD). a-Synuclein was
the first protein linked to familial PD.
Understanding the biology of a-synuclein
attracted extensive attention because it
is the main protein that accumulates in
Lewy bodies, which are the pathological
hallmark of sporadic PD. The involvement of
a-synuclein in both familial PD and sporadic
PD provides a critical linkage between
familial PD and sporadic PD. Research into
the pathophysiology of a-synuclein indicates
that this protein tends to aggregate, and
that the mutations associated with familial
disease increase this tendency to aggregate.
The tendency of a-synuclein to aggregate
presents a common theme that is apparent
in many neurodegenerative diseases
because many of the pathogenic proteins
tend to aggregate. Aggregation, or more
specifically, oligomerization and fibrillization,
are essential steps in toxicity related to
both Ab and tau toxicity. Four other genes
have been clearly associated with familial
Parkinson’s disease, and more genes carrying
mutations associated with familial PD are
likely to be identified. Loss of functions
mutations in parkin, DJ-1 and Pink-1 all
cause familial PD, while mutations in LRRK2
associated with familial PD are thought to
cause a gain of function. Parkin is an E3-
ubiquitin ligase; LRRK2 binds parkin. The
role of E3-ligases in proteasomal function
suggests that proteasomal dysfunction
could contribute to familial PD. On the other
hand, toxicological studies indicate that
chemicals inhibiting mitochondrial function,
such as MPTP or rotenone, can mimic
some aspects of PD. Pink-1, DJ-1, LRRK2
and parkin all associate with mitochondrial
structures, and neurodegeneration caused
by over-expressing a-synuclein also
causes mitochondrial degeneration. These
studies present tantalizing clues about
the causes of PD, but a comprehensive
understanding of the mechanism of
degeneration remains elusive.
Disorders of the Nervous System: New Tools to Examine New Pathways
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Polyglutamine-associated neurodegenerative disorders: Expansions of polyglutamine-repeat
domains are associated with 7 different
diseases, including Huntington’s disease,
Spinobulbar muscular atrophy, 6 forms of
spinocerebellar ataxias and dentatorubral-
palidoluysian atrophy. Proteins harboring
expanded polyglutamine repeats aggregate
readily. The aggregates accumulate in the
nucleus or the cytoplasm, depending on the
cell type. Aggregation in the cytoplasm or
endoplasmic reticulum causes proteasomal
inhibition. Nuclear aggregation is associated
with transcriptional changes, possibly
because some transcription factors, such
as CREB, contain polyglutamine repeat
domains and tend to be sequestered by the
nuclear aggregates. Studies using animal
models suggest that loss of CREB function in
polyglutamine repeat disorders contributes
to the neuronal dysfunction. Treatment
with HDAC inhibitors counteracts the loss
of CREB function and is now being tested
in patients with Huntington’s disease.1
Amyotrophic Lateral Sclerosis: The
mechanisms producing amyotrophic lateral
sclerosis (ALS) remain enigmatic. ALS is an
uncommon disease. Familial ALS, which
accounts for about 10% of the cases,
is caused by mutations in superoxide
dismutase. Although the mutations increase
the tendency of superoxide dismutase
to aggregate, the mutations appear to
cause disease by altering the activity of
superoxide dismutase, perhaps by causing
release of the copper or zinc at its core,
rather than by a mechanism linked to
aggregation. The relationship between
familial and sporadic ALS, though, remains
elusive. Sporadic ALS might be linked
to dysfunction of glutamate uptake by
glia, but this theory is controversial.
Different diseases converging onto common pathways
Understanding why each particular
biochemical abnormality is deleterious to
the brain represents one of the central goals
of research for many of these diseases. The
biochemical dysfunction that underlies each
disease induces a number of processes that
are detrimental to the cell including produc-
tion of reactive oxygen species, activation
of stress kinases and ultimately activa-
tion of the apoptotic signaling cascade.
These processes are also stimulated during
the types of acute injury that occur with
trauma, stroke, status epilepticus and
binging on addictive drugs. Despite a wide
variety of different insults that initiate the
neuronal injury, the brain shows cellular
responses that generalize across a broad
number of diseases.
The stress kinase cascade flows through
a large number of MAP kinases and
MAPKAP kinases to converge on a number
of terminal kinases that include the c-Jun
kinase and p38 family of stress kinases.
DNA damage activates repair systems
through enzyme cascades mediated by p53
and other checkpoint proteins. The p53
protein also regulates a large number of
cellular and mitochondrial responses. Small
gaseous signaling agents, such as nitric
oxide and carbon monoxide, modulate
signaling cascades that can be good or bad,
depending on the cell type. The apoptotic
cascade flows to and then emanates from
the mitochondria. This cascade proceeds
via a number of linked biochemical reac-
tions including caspase activation, reaction
by the Bcl-2 family of proteins, release of
cytochrome C, and modulation of other
proteins that regulate apoptosis such as IAP
and Diablo. The mitochondria respond to
the damage, producing free radicals due
to electron leakage and reverse electron
transport. The production of free radicals
combined with changes in calcium content
and other signals act to open the perme-
ability transition pore. Increased calcium
also causes damage by activating calcium
dependent proteases, and the energy
demands that are sometimes associated
with excitotoxicity put further strain on the
capacity of the mitochondria, driving down
cellular energy stores.
Extrinsic signaling also modifies neurode-
generation. Degeneration in the CNS is
frequently accompanied by an activated
inflammatory response, which is presum-
ably designed to remove the damaged
tissues. Inappropriate activation of immune
responses in the brain lies at the heart of
multiple sclerosis. The cytokines and reac-
tive oxygen species (ROS) produced by the
microglia are directly toxic to oligodendro-
cytes and neurons; production of cytokines
and ROS plays an important role acceler-
ating degeneration in AD, PD and ALS.
Understanding how to dampen or direct
the immune response represents one of
the many challenges facing researchers of
neurodegeneration.
The degenerative processes can be counter-
acted by growth factors, which activate
signal transduction cascades that promote
survival and cell growth. These cascades
induce a reactive response that stimulates
synaptic plasticity at the same time as other
elements of the neurons are undergoing
degeneration. Many studies have shown
that factors that stimulate pro-survival
cascades delay degeneration. Brain derived
nerve growth factor, nerve growth factor
and glial derived growth factor all have
been shown to be neuroprotective in
cellular and animal models.
The brain also has the capacity to respond
to the neuronal injury by producing new
neurons in some brain areas from stem
cells. These pluripotent cells can replace lost
neurons, and ultimately might represent an
endogenous source of cellular replenish-
ment for the brain. The cytokines and ROS
produced by the disease process, though,
appear to harm stem cell production in
addition to inducing neurodegeneration.
Many studies show that the capacity to
produce stem cells is reduced in animal
models of neurodegenerative disease.
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Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Review Article on the Field of Neuroscience (continued)
Each of these factors – signaling, growth
factors and regeneration – impinge on
the neuron to modulate synaptic function.
Increases in our understanding of synaptic
biology have lead to dramatic advances
in our understanding of the biochemical
mechanisms underlying diseases such as
schizophrenia, depression, drug addic-
tion and autism. The synapse is a finely
tuned microscopic engine not dissimilar
to a nano-robot. It has numerous inter-
connected elements that must respond
in concert to achieve the optimal level
of neuronal signaling. Subtle changes in
the amount or type of signaling lead to
dramatic changes in disorders that manifest
in altered behavior. Families of proteins,
such as neuregulins and dysbindin, modu-
late activity of the glutamate receptors.
Mutations at different points in this regula-
tory cascade appear able to predispose to
schizophrenia when environmental insults
are also present to stress the system. For
instance, mutations in neuregulin I type 3
cause dysfunction of glutamate receptors
and cholinergic neurons and appear to be
linked to schizophrenia.
Serotonin neurotransmission plays a central
role in depression, with low serotonin
precipitating depression. Medications, such
as the selective serotonin reuptake inhibi-
tors (SSRIs), can alleviate the depression
but the slow kinetics of the response of
depressed patients to SSRIs suggests that
other processes need to occur in parallel.
SSRIs are known to stimulate growth factor
responses and also have been recently
shown to slowly modulate a protein termed
P10. These processes provide important
clues about the pathophysiology of depres-
sion. Activity of dopaminergic neurons is
associated with drug abuse, but too much
activity stimulates glutatamate-mediated
neurotoxicity. In addition, the overwhelming
pleasure stimulated by drugs of abuse and
the resulting craving produce incapacitating
behavioral responses. Medications that
reduce the strength of the response, such
as naltrexone, show promise for treatment
of some addictions, but understanding how
to intervene in this circuitry remains a major
challenge in the field of behavioral neurosci-
ence. Our increasing understanding of the
cell biology of the addictive axis provides
a corresponding increase in the number of
strategies and targets for intervention.
High throughput technologies: Arrays, multiplexing and systems biology
Disease processes are typically complex.
New technology allows the investigator to
characterize multiple responses occurring
in parallel. High throughput technology
has emerged as a key approach towards
dissecting the complex series of biochemical
events that occur in parallel in the brain.
This technology can provide a broad over-
view of the magnitude and kinetics of
changes in gene expression, protein expres-
sion or post-translational modifications.
Gene arrays provide an overview of the
broad spectrum of transcriptional responses
to any particular signal. Three different
technologies are also emerging to provide
overviews of the response of the proteome
in stressed or injured tissues. 1) Antibody
arrays allow analysis of how the levels of
hundreds or thousands of proteins respond
to a stress, such as treatment with Ab. 2)
Two-dimensional gel technology combined
with mass spectroscopy also allows identifi-
cation of proteins that might not be picked
up through antibody arrays. 3) Multiplexing
provides accurate quantification of dozens
of targets in one rapid assay.
Multiplexing is the term used to describe
assays that measure multiple epitopes
at once. Bead technology allows parallel
analyses of multiple epitopes in one assay
using either dedicated multiplexing devices
or BD FACS analyzers, which have the
benefit of being available to many labora-
tories through institutional core facilities.
Multiplexing allows any laboratory to
affordably profile metabolic responses of
tissues. Multiplexing technology is also
highly valuable for situations where the
tissues are available in only limited
quantities. By analyzing multiple proteins in
parallel in the same sample, researchers are
able to maximize the information gained
from small amounts of sample. The ability
to dissect, categorize and quantify protein
networks in tissues is increasingly being
applied to facilitate translational research
in an effort to move today’s breakthroughs
into tomorrow’s medicines.
Reagents are now available to identify and
quantify the cascades of protein activity
associated with stimulation of biochemical
pathways. Antibodies specific to phosphory-
lated proteins allow determination of the
amount of activation of a particular protein
and the kinetics of activation. Patterns of
activity of proteins can be studied for indi-
vidual proteins or for groups of proteins.
The latter analysis allows comparison of
activities of different pathways or validation
of activation by analysis of multiple steps
in a particular signal transduction pathway.
The commercial sector offers strong libraries
of reagents to facilitate studies of signal
transduction.
Imaging: The science of real estate
Accumulating evidence demonstrates the
immense importance of location in neuro-
biology. The delineation of whether an
inclusion occurs in the nucleus, cytoplasm,
mitochondria or extracellular domain has
important implications for mechanisms
of degeneration and potential avenues of
therapy. Identifying which synapses are
subject to oxidative damage or post-trans-
lational modification focuses attention on
particular neuronal cell types and particular
receptor interactions. Determining whether
inflammation targets a neuron, a glia or
an oligodendrocyte helps to define the
disease process. The steady march of cell
culture methods facilitates our knowledge
of neurobiology by allowing researchers
to analyze isolated groups of cells, but
novel imaging technologies, novel antibody
probes and novel fluorophores greatly
expand our imaging capabilities. Multiple
excellent fluorophores are now available to
bdbiosciences.com9Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
facilitate double, triple and even quadruple
staining protocols. Confocal microscopy
allows 3-dimensional reconstructions of
molecular distributions within a neuron or
glial cells. Using confocal microscopy we
can determine whether epitopes are actually
co-localized or simply vertically juxtaposed.
Imaging technologies such as fluorescence
resonance transfer can test whether puta-
tive molecular interactions actually occur
in living cells. Imaging has also moved in
vivo. Magnetic resonance imaging and
positron emission tomography has been
used for years to probe the brain, but now
techniques such as two-photon imaging
allows analysis of cellular events in the living
brain. Developing imaging technology and
improved biomarkers offers the ability to
study structures such as amyloid plaques
and neurofibrillary tangles in living animals.
The future of disease research is likely
to shift towards more complex analyses
as systems biology is applied to disease
processes. Massive parallel studies of gene
arrays and protein interaction screens are
already revolutionizing our understanding
of the genetic events that take place in
a developing nematode or proliferating
bacteria. This type of technology will
increasingly be applied to understand the
human body, both in health and in disease.
Multiplexing will need to be applied to
bridge the gap between gene arrays,
systems biology and disease research.
Identifying key signaling pathways and
quantifying the selective changes that lead
to particular diseases will certainly facilitate
developing diagnostic tests and therapeutic
treatments for the diseases that plague our
populations today.
bdbiosciences.com10 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Cell Culture
Applying Innovative Technology to Neuronal Cell Culture
As the first company to produce sterile,
disposable labware more than 50 years
ago, BD Biosciences is a worldwide leader
in top-quality cell culture products. Our
comprehensive line of innovative products
will meet your neuronal cell culture needs.
Consistent cell culture conditions are
required for reproducible research results.
For many cell lines, the easiest solution is
to use a treated surface that allows cells
to attach. The consistency of this surface
depends on the treatment method used.
Many manufacturers have long used atmos-
pheric plasma treatments (eg, corona) to
create hydrophilic surfaces. In corona treat-
ment, a high-voltage discharge creates
a reactive gas plasma above the growth
surface of the vessel. In this process, the
highly interactive gas plasma mixture is
exposed to ambient air. The consistency
of the treatment surface can therefore be
compromised by day-to-day environmental
changes.
At BD Biosciences, molded polystyrene
vessels are placed in a closed, highly-
controlled environment that ensures a
consistent treatment surface.2,3
BD Biosciences made a major research
investment to develop this unique vacuum-
gas plasma process, which
results in a hydrophillic surface on the
plastic that supports cell attachment and
spreading.4-6
This process is used to produce both
traditional tissue culture (TC) surfaces on
BD Falcon™ dishes, plates, flasks, and roller
bottles, as well as surfaces on BD Primaria™
cultureware. Recent research published in
Stem Cells demonstrates the superiority of
the BD Biosciences process.3
Cell performance tests ensure consistent results
A sensitive clonogenic assay85 using MRC-5
cells, a diploid human fibroblast line, is used
to validate the manufacturing process for
each BD Falcon TC product. Routine testing
of standard TC products is performed by
testing growth to confluency at 72 hours
with MRC-5 cells. The surface chemistry
of each lot of BD Primaria products is
confirmed by electron spectroscopy for
chemical analysis (ESCA).
Our Commitment to Quality
All BD Falcon, BD BioCoat, and
BD™ brand product manufacturing
sites are ISO 9000 certified. This
certification verifies that our facilities
meet international quality system
standards. The quality system is
routinely audited by a notified body
to ensure a work environment that
consistently maintains the highest
standards. ISO compliance gives our
customers an added level of assurance
that BD Biosciences is committed
to superior quality and continuous
product improvement. Copies of our
certificates can be obtained by calling
your local BD office.
Advanced Culture Surfaces for Neuronal Cells
Pseudo-colored merged image of Alexa Fluor® 488 (cell bodies and neurites: green) and Hoechst (nuclei: blue) channels of PC12 cells differentiated with 100 ng/mL NGF. Images were acquired as a 4x4 montage using a 20x objective (0.75 NA).
bdbiosciences.com11Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
BD Falcon Flasks
description qty./pack qty./case cat.no.
CELLineTM 1000 System 1 3 353137
BD Falcon 75 cm2 standard TC Flask, vented cap 5 60 353136
BD Falcon 150 cm2 standard TC Flask, plug-seal cap 5 40 355000
BD Falcon 150 cm2 standard TC Flask, vented cap 5 40 355001
BD Falcon 175 cm2 standard TC Flask, vented cap 5 40 353112
BD Falcon Dishes
description qty./pack qty./case cat.no.
BD Falcon 35 mm Easy-Grip Style 20 500 351008Bacteriological Petri Dish
BD Falcon 35 mm Easy-Grip Style Cell Culture Dish 20 500 353001
BD Falcon 60 mm Center-Well Organ Cell Culture Dish 20 500 353037
BD Falcon 100 mm Standard Style Cell Culture Dish 20 500 353003
BD Falcon 150 mm Integrid Dish 10 100 351013
BD Falcon Cell Culture Plates
description qty./pack qty./case cat.no.
BD Falcon 6-well Multiwell Plate, flat-bottom 6 36 353224
BD Falcon 12-well Multiwell Plate, flat-bottom 1 50 353043
BD Falcon 24-well Multiwell Plate, flat-bottom 6 36 353226
BD Primaria 24-well Multiwell Plate, flat-bottom 1 50 353847
BD Falcon 48-well Multiwell Plate, flat-bottom 6 36 353230
BD Falcon 96-well Microplates, TC, round-bottom 1 50 353077
BD Falcon 96-well Microplates, TC, flat-bottom 1 50 353070
BD Falcon 96-well Microplates, Standard, flat-bottom 1 50 351172
BD Falcon 96-well Microplates, Standard, round-bottom 1 50 351177
BD Falcon 384-well Optilux™ Microplates 5 50 353962
BD Falcon 1536-well Microplates 15 60 353249
BD Falcon Flasks
BD Falcon™ flasks are used to passage or to scale-up cells for
production. Vacuum-gas plasma tissue culture treatment provides
consistent cell attachment, spreading, and growth. Choose BD
Primaria™ non-treated polystyrene for standard tissue cultures or
BD BioCoat™ growth surfaces to meet your individual cell culture
requirements.
Visit bdbiosciences .com/flasks for a complete listing of BD Falcon flasks.
BD Falcon Cell Culture Plates
BD Falcon cell culture plates are available in well-characterized
dimensions and surface chemistries. These properties allow the use
of divergent cell culture methodologies and can be used for fluores-
cence, luminescence, colorimetric, and radiometric assays. Available
in configurations from 6- to 1536-well formats, these plates provide
the flexibility to meet the needs of a wide range of cell-based assays
and other biochemical assay needs in both manual and automated
settings.
BD Falcon Dishes
BD Falcon dishes are available in a wide range of sizes and
configurations to ensure they meet the needs of more
specialized applications.
The BD Falcon™ cultureware family also includes cell scrapers for
removing attachment cells from cultureware, cell strainers for
enhancing cell isolations from tissues, and various storage and
collection containers. Visit bdbiosciences .com/cellcultureware for
more information on these products.
Visit bdbiosciences .com/cellcultureplates for a complete listing of BD Falcon cell culture plates.
Visit bdbiosciences .com/cellculturedishes for a complete listing of BD Falcon cell culture dishes. Visit bdbiosciences .com/specialtydishes for more information on our bacteriological, compartment, and other specialty dishes.
BD Falcon Cultureware
bdbiosciences.com12 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
BD Falcon Tubes
description qty./pack qty./case cat.no.
BD Falcon 50 mL Conical Tubes 25/bag 500 352070
25/rack 500 352098
BD Falcon 15 mL Conical Tubes 125/bag 500 352096
50/rack 500 352097
BD Falcon 5 mL Round-Bottom Tubes 125/pack 1000 352052
BD Falcon 8 mL Round-Bottom Tubes 125/pack 1000 352027
BD Falcon 14 mL Round-Bottom Tubes 25/pack 500 352059
25/pack 500 352057
BD Falcon Pipets
description qty./pack qty./case cat.no.
BD Falcon 1 mL Serological Pipets 25/bag 1000 357506
100/box 1000 357521
BD Falcon 5 mL Serological Pipets 50/bag 200 357543
BD Falcon 10 mL Serological Pipets 50/bag 200 357551
BD Falcon 25 mL Serological Pipets 50/bag 200 357525
BD Falcon 50 mL Serological Pipets 25/bag 100 357550
BD Falcon Pipets
BD Falcon™ pipets, first in quality and first in innovation, are
manufactured at our ISO 9001-certified facility in a state-of-the-art,
high-performance work environment. Dedicated product teams use
advanced technologies and techniques, combined with rigorous
quality control procedures, to ensure the integrity of every pipet.
Each pipet component is made of the highest quality polystyrene
resin. Process control and attention to detail guarantee the finest
quality product. A critical volumetrics test verifies the accuracy of
each pipet by measuring the delivered volume. The final result is
consistently high-quality pipets that provide precise and accurate
liquid handling.
BD Falcon Tubes
Your life science research demands the most stable and controlled
environment possible for the analysis of biological and chemical
samples. At BD Biosciences, we manufacture our BD Falcon™
conical and round-bottom tubes from advanced bioanalytical-grade
resins. Our polymer selection process includes extensive testing to
ensure that the polymer does not leach unwanted substances.
BD Falcon tubes and our unique medical-style packaging provide
unsurpassed convenience and consistency.
BD Falcon Pipets and Tubes
Visit bdbiosciences .com/pipets for a complete listing of BD Falcon pipets.Visit bdbiosciences .com/tubes for a complete listing of BD Falcon tubes.
Cell Culture (continued)
bdbiosciences.com13Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Unique surface chemistry for enhanced cell culture
BD Biosciences has made a significant
research investment to develop a unique
vacuum-gas plasma process used to
produce both BD Primaria and traditional
tissue culture (TC) surfaces on BD Falcon
dishes, plates, flasks, and roller bottles.
The incorporation of nitrogen-containing
cations has been correlated to the attach-
ment and spreading of primary endothelial
cells in a clonal cell-growth assay.2 The
gases used to manufacture BD Primaria
cultureware contain both oxygen and
ammonia, resulting in the incorporation of
a variety of nitrogen-containing functional
groups on the surface, in addition to the
negatively charged oxygen-containing
groups found on traditional TC surfaces.
Because the complex surface on
BD Primaria cultureware is homogeneous
and stable, it has been used by researchers
for over a decade to improve attachment
and differentiation of a variety of cell types.
For example, cell biologists have used
BD Primaria cultureware for cultivating
neuronal cells,7 hepatocytes,8,9 and endo-
thelial cells.10 The surface chemistry of BD
Primaria products is confirmed by electron
spectroscopy for chemical analysis (ESCA).
BD Primaria Cultureware
When chick embryo spinal cord neurons are cultured on BD Primaria cultureware (left), growth is enhanced and extensive neurite development occurs. In this experiment, cells clumped and detached from traditional TC plates (above) after 20 days in culture, but remained viable and differentiated on BD Primaria cultureware.
Note: At pH 7, carboxy groups may be slightly dissociated and assume a negative (anionic) charge. Amine groups may protonate and assume a positive charge (cationic).
Traditional Tissue Culture Surface Chemistry BD Primaria™ Surface Chemistry
35 x 10 mm Easy-Grip Dish
description qty./pack qty./case cat.no.
Actual Dimensions: 40.28 mm O.D. x 6.17 mm
Actual Growth Area: 11.78 cm2
Working Volume: 2.5-3.0 mL 20 200 353801
60 x 15 mm Standard Dish
description qty./pack qty./case cat.no.
Actual Dimensions: 54.81 mm O.D. x 13.26 mm
Actual Growth Area: 21.29 cm2
Working Volume: 6.0-7.0 mL 20 200 353802
100 x 20 mm Standard Dish
description qty./pack qty./case cat.no.
Actual Dimensions: 89.43 mm O.D. x 19.18 mm
Actual Growth Area: 58.95 cm2
Working Volume: 16.0-17.5 mL 20 200 353803
25 cm2 Flask with Canted Neck
description qty./pack qty./case cat.no.
Nominal Growth Area: 25 cm2
Total Volume: 50 mL
Plug-Seal Caps 20 200 353813
Vented, 0.2 µm Caps 20 100 353808
75 cm2 Flask with Straight Neck
description qty./pack qty./case cat.no.
Nominal Growth Area: 75 cm2
Total Volume: 250 mL
Plug-Seal Caps 5 100 353824
Vented, 0.2 µm Caps 5 100 353810
6-well Multiwell Plate with Lid
description qty./pack qty./case cat.no.
Growth Area: 9.6 cm2
Working Volume: 15.5 mL 1 50 353846
24-well Multiwell Plate with Lid
description qty./pack qty./case cat.no.
Growth Area: 2.0 cm2
Working Volume: 3.5 mL 1 50 353847
96-well Multiwell Plate Flat-bottom with Lid
description qty./pack qty./case cat.no.
Growth Area: 0.32 cm2
Working Volume: 0.37 mL 1 50 353872
bdbiosciences.com14 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
BD Primaria™ Cultureware (continued)
Cell Culture (continued)
bdbiosciences.com15Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Extracellular Matrix Proteins and Attachment Factors
description qty. cat.no.
BD™ Extracellular Matrix, human 1mg 354237
BD Cell-Tak™ Cell and Tissue Adhesive 1mg 354240
5mg 354241
(2x5mg)10mg 354242
BD™ Osteopontin 50µg 354256
BD Collagen I, bovine 30mg 354231
BD Collagen I, human 250µg 354243
BD FIBROGEN Collagen I, human recombinant 250µg 354254
BD Collagen I, rat tail 100mg 354236
(10x100mg)1g 356236
BD Collagen I High Concentration (HC), rat tail 100mg 354249
BD Collagen II, bovine 5mg 354257
BD Collagen III, human 250µg 354244
BD FIBROGEN Collagen III, human recombinant 250µg 354255
BD Collagen IV, human 250µg 354245
BD Collagen IV, mouse 1mg 354233
(10x100µg)1mg 356233
BD Collagen V, human 250µg 354246
BD Collagen VI, human 500µg 354261
BD Fibronectin, human 1mg 354008
5mg 356008
(5x500mg)25mg 356009
BD Laminin, mouse 1mg 354232
BD Laminin/Entactin Complex High Concentration, mouse 10.5mg 354259
BD Ultrapure Laminin, mouse (entactin-free) 1mg 354239
BD Poly-D-Lysine 2mg 354210
BD Vitronectin, human 250µg 354238
FIBROGEN Human Recombinant Collagens are manufactured for BD Biosciences by FibroGen, Inc.
Visit bdbiosciences .com/ecms for more product information and technical references.
The development and normal functioning of cells depends on inter-actions with molecules in their microenvironment. The major classes of molecules that regulate cellular development and function include growth and differentiation factors, cell adhesion molecules, and the components of the extracellular matrix (ECM).
The ECM is a vital component of the cellular microenvironment in vivo. It serves as the framework for cell anchorage and plays a dynamic role in the regulation of cell growth and function.
The ECM exerts influences on behavior (adherence, spreading, differentiation, and migration) and the pattern of gene expression of the cells in contact with it. The ECM is not static but changes during both normal development and in tissue repair and regeneration; thus, is intimately involved in both normal biological function and response to injury.4
To create physiologically relevant in vitro models that support normal cell growth and function, the components of the in vivo environ-ment must be incorporated. Use of ECM as a coating for TC surfaces permits the development of model systems that closely mimic in vivo conditions.
Although the ECM of different tissues is specialized, the components commonly include collagens, fibronectin, laminins, proteoglycans, and vitronectin. Purified ECM proteins can be used in in vitro culture
systems to provide an appropriate environment for specific cell types and applications. The presence of ECM components promotes cell attachment, spreading, growth, and differentiation of a variety of primary cells and cell lines including neuronal stem cells. For some cells, the presence of particular ECM components is critical for optimum cell growth in vitro. Purified ECM proteins can be easily used to coat the desired tissue culture surface. Alternatively, BD BioCoat™ cellware, including tissue culture plates, dishes, flasks, coverslips, and cultureslides, are available pre-coated with a variety of ECM proteins and attachment factors. See additional BD BioCoat Cellware on pages 18–28.
Choice of ECM Substrata In Vitro
Nerve cells are more selective in their choice of substrata than most other cell types. Selection of an optimal substratum for neuronal cells in culture is often an empirical process. Primary neuronal cells exhibit minimal to no growth or differentiation on untreated glass or plastic surfaces. The choice of ECM or surface modification is an important component to consider when optimizing neuronal cell cultures. Recognizing the increasingly important role the ECM plays in the regulation of fundamental cellular processes, BD Biosciences offers a wide range of ECM proteins and attachment factors for researchers to incorporate into their cell culture systems.
Extracellular Matrix Proteins and Attachment Factors
*NA - not applicable †ND - not determined
bdbiosciences.com16 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
• Promotes differentiation of many cell types
• Gels to form a three-dimensional model of basement membrane
• Increases rate of human tumor cell growth in nude mice
• Creates models to measure invasive potential of tumor cells
BD Matrigel™ basement membrane matrix
is a solubilized basement membrane
preparation extracted from the Engelbreth-
Holm-Swarm (EHS) mouse sarcoma, a tumor
rich in ECM proteins. Its major component
is laminin, followed by collagen IV, entactin,
and heparan sulfate proteoglycan.87-90 It also
contains TGF-b, fibroblast growth factor,
tissue plasminogen activator,91 and other
growth factors that occur naturally in the
EHS tumor.
At room temperature, BD Matrigel Matrix
polymerizes to produce a biologically active
matrix material resembling the mammalian
cellular basement membrane.
BD Matrigel Matrix is effective for the
attachment and differentiation of both
normal and transformed anchorage-
dependent cell types including neurons,
epithelial, and oligodendrocytes. These
include neurons, hepatocytes, Sertoli cells,
mammary epithelial cells, melanoma cells,
vascular endothelial cells, thyroid cells,
and hair folicle cells.88,92,94,95 BD Matrigel
basement membrane matrix will influence
casein gene expression in mouse mammary
epithelial cells.93,96 It also supports in vivo
peripheral nerve regeneration89 and
facilitates differentiation of bovine oviduct
epithelial cells.97
Growth Factor Reduced (GFR) BD Matrigel
Matrix is useful as an alternative to BD
Matrigel Matrix in many of the applications
listed above when a more highly defined
basement preparation is desired. It has
been used to define the signals necessary
in the formation of canicular cell processes
in bone cells,98 in elucidating the role of
growth factors in the formation of tubules
by primary mouse kidney cells,86 and in
gene expression studies of primary mouse
mammary epithelial cells.99 This phenol
red-free product is suitable for those assays
that require color detection (eg, colorimetric
assays and fluorescence detection).
All BD Matrigel basement membrane
matrices, including the High Concentration
formulation, can also be used to assess
in vivo angiogenic activity of different
compounds by subcutaneous injection
into mice (BD Matrigel Plug Assay).100-104
A similar approach can be used for the
transplantation and investigation of human
tumor cells. Reports include studies with
prostatic, breast, small-cell lung, colon,
adrenal carcinomas, melanomas, and
lymphoblastic leukemia cells.105-110
BD Matrigel Basement Membrane Matrix
Human Submandibular Gland (HSG) cells cultured on GFR BD Matrigel™ Matrix differentiate to form acinar structures within 24 hours. The acini shown were stained with H/E at 72 hours. Indirect immunofluorescence staining also revealed the salivary gland specific cysteine protease inhibitor, cystatin, in HSG cell acini (data not shown). (Photo courtesy of Dr. Hynda Kleinman.)
Amounts of Growth Factors (GF) Present in BD Matrigel™ Matrix vs. GFR BD Matrigel™ Matrix
Growth factor
Range of GF Concentration in
BD Matrigel MatrixAverage GF Concentration
in BD Matrigel MatrixTypical GF Concentration
in GFR BD Matrigel Matrix
EGF 0.5–1.3ng/mL 0.7ng/mL <0.5ng/mL
bFGF <0.1–0.2pg/mL NA* ND†
NGF <0.2ng/mL NA* <0.2ng/mL
PDGF 5–48pg/mL 12pg/mL <5pg/mL
IGF-1 11–24ng/mL 16ng/mL 5ng/mL
TGF-b 1.7–4.7ng/mL 2.3ng/mL 1.7ng/mL
Cell Culture (continued)
BD Matrigel™ Basement Membrane Matrix
description qty. cat.no.
Formulation: Dulbecco’s Modified Eagle’s Medium with 10 µg/mL gentamycin. Typical protein concentrations are between 9-12 mg/mL. BD Matrigel Matrix is compatible with all culture media.
5mL 356234
10mL 354234
(5x10mL)50mL 356235
BD Matrigel Matrix High Concentration (HC)
description qty. cat.no.
Formulation: Dulbecco´s Modified Eagle´s Medium with 10 µg/mL gentamycin. Typical protein concentrations are between 18-22 mg/mL. BD Matrigel Matrix HC is compatible with all culture media.
10mL 354248
BD Matrigel Matrix, Phenol Red-free
description qty. cat.no.
Formulation: Dulbecco’s Modified Eagle’s Medium (without phenol red) with 50 µg/mL gentamycin. Phenol Red-free BD Matrigel Matrix is compatible with all culture media.
Standard Concentration 10mL 356237
High Concentration 10mL 354262
Growth Factor Reduced (GFR) BD Matrigel Matrix
description qty. cat.no.
Formulation: Dulbecco’s Modified Eagle’s Medium with 50 µg/mL gentamycin. GFR BD Matrigel™ Matrix is compatible with all culture media.
Purification: Purified by the method of Taub, et al., reducing the level of heparan sul-fate proteoglycan and several growth factors (eg, EGF, bFGF, IGF-1, PDGF, and NGF, but not TGF-b).86
Standard Concentration 5mL 356230
Standard Concentration 10mL 354230
High Concentration 10mL 354263
GFR BD Matrigel Matrix, Phenol Red-free
description qty. cat.no.
Formulation: Dulbecco’s Modified Eagle’s Medium (without phenol red) with 50 µg/mL gentamycin. Phenol Red-free BD Matrigel Matrix is compatible with all culture media.
Purification: Purified by the method of Taub, et al., reducing the level of heparan sul-fate proteoglycan and several growth factors (eg, EGF, bFGF, IGF-1, PDGF, and NGF, but not TGF-b). 86
10mL 356231
Dispase
Dispase is a bacillus-derived neutral metalloprotease that is recom-
mended for recovering cells cultured on BD Matrigel basement
membrane matrix. BD™ Dispase will yield a single cell suspension
far more gently and effectively than trypsin, collagenase, or other
proteolytic enzymes; it will not harm cells harvested for subcultiva-
tion or bioassays. In addition, BD Dispase may be used for tissue
dissociation. BD Dispase cleaves fibronectin, collagen IV, and to a
lesser extent collagen I, but it does not cleave collagen V or laminin.
Cell Recovery Solution
BD™ Cell Recovery Solution allows for the recovery of cells cultured
on BD Matrigel Basement Membrane Matrix for subsequent
biochemical analyses. BD Cell Recovery Solution depolymerizes
BD Matrigel Matrix gels without enzymatic digests and lengthy
incubation periods at high temperatures. Cells are released without
damage, thereby avoiding biochemical changes during incubation
and digestion of extracellular portions of cell-surface receptors and
adhesion molecules.
Cell Recovery Solution
description qty. cat.no.
BD Cell Recovery Solution 100mL 354253
BD Dispase
description qty. cat.no.
BD Dispase (5,000) caseinolytic units 100mL 354235
ECM Composition of BD Matrigel Matrix vs GFR BD Matrigel Matrix
Basement MembraneComponent
Percent in BD Matrigel Matrix
Percent inGFR BD Matrigel Matrix
Laminin 56% 61%
CollagenIV 31% 30%
Enactin 8% 7%
bdbiosciences.com17Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
bdbiosciences.com18 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Pre-coated for Reproducibility and Convenience
BD BioCoat cellware combines the quality
of BD Falcon TC plates with a variety of
ECM proteins and attachment factors. The
result promotes cell attachment, spreading,
growth, and differentiation of a variety of
primary cells and cell lines in serum-free or
serum-containing cultures. A proprietary
manufacturing process applies the ECM
components to the vessel surface. The result
is a uniform, optically clear matrix substrate.
This technology, together with our exacting
quality control, guarantees the performance
of each lot, as well as consistency
from lot to lot.
Applications include:• Cell growth
• Cell adhesion assays
• Receptor-ligand binding assays
• Routine drug screening assays
• Studies of tissue morphogenesis
• Studies of cell-matrix interactions
• Regulation of signal transduction and gene expression
BD BioCoat Advantages
Ready-to-Use Convenience
Spend more time performing your
experiments rather than preparing for
them. Pre-coated BD BioCoat cellware
saves time and labor costs while
increasing productivity.
Quality Assurance Testing
Each lot of BD BioCoat cellware is
thoroughly tested for bioactivity and guar-
anteed to perform as claimed so you can
use them with complete confidence.
Reliable Performance
BD BioCoat cellware improves cell attach-
ment and increases proliferation rates for a
variety of normal and transformed cells.
Lot-to-Lot Consistency
BD Biosciences prides itself on maintaining
highly controlled ISO 9001 production
environments and validated manufacturing
procedures that result in uniformity and
consistent performance.
Wide Selection
Available with a wide range of ECM
proteins and attachment factors,
BD BioCoat cellware helps optimize
conditions for attachment, growth, or
differentiation for your cell type.
Custom Coatings
BD BioCoat Custom Coating Service
offers an extensive selection of cell-based
coatings on a wide variety of BD Falcon
vessels to meet your specialized needs.
BD BioCoat Cellware
Cell Culture (continued)
bdbiosciences.com19Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Flasks are available in various sizes and designs with
standard, plug-seal, and vented cap options.
Multiwell plates are manufactured using a crystal-
grade polystyrene and feature patented labyrinth lid,
condensation rings, and deep-well design to control
contamination while reducing evaporation and mini-
mizing edge effects.
Dishes are exceptionally flat for distortion-free
optics and feature stacking rings for easier stacking
and handling.
CultureSlides have an innovative sealing design that
minimizes leakage and a plastic chamber affixed to a
specially cleaned glass slide that can be removed with
an easy-to-use, disposable safety removal tool.
BD BioCoat coverslips are composed of No.1 German
glass and provide an optically clear surface that is
non-neurotoxic and exhibits low background fluores-
cence. The convenient package also acts as a storage
container and allows for easy coverslip manipulation.
BD BioCoat coverslip-bottom dishes are 35 mm
dishes with a coverslip bottom that are easy to use
and facilitate preparation of cells for microscopic
analysis. The coverslip floor is No.1 German glass.
This format is ideal for use in high resolution and
inverted microscopy, fluorescence imaging in live
cells, confocal microscopy, phase contrast microscopy,
and micromanipulations.
BD Matrigel basement membrane matrix
is a solubilized basement membrane
preparation extracted from the Engelbreth-
Holm-Swarm (EHS) mouse sarcoma, a tumor
rich in ECM proteins. Its major component
is laminin, followed by collagen IV, heparan
sulfate proteoglycans, entactin, and
nidogen. BD Matrigel Matrix is effective for
the attachment and differentiation of both
normal and transformed anchorage depen-
dent cell types including neurons, epithelial,
and oligodendrocytes.
BD BioCoat Matrigel Matrix Cellware applications include:
• Elicitation of tissue-specific cellular morphology and protein production in epithelial cells
• Differentiation of neuronal, endothelial, and muscle cells
• Development of three-dimensional matrix model systems
BD BioCoat Matrigel Matrix has been used to culture:
• Rat hepatocytes26
• Primary human hepatocytes27
• Mouse pituitary gland tissue28
• Rabbit colonocytes29
• Human urothelial cells30
• Osteopontin (OPN) deficient rat vascular smooth muscle cells31
Source:• EHS mouse tumor
Formulation:• Dulbecco’s Modified Eagle’s Medium
with 50 μg/mL gentamycin
• BD Matrigel Matrix is compatible with all culture media
Quality Control:• Tested for ability to promote neurite
outgrowth from chick dorsal root ganglia in the absence of NGF
• Tested and found negative for bacteria and fungi
Storage and Stability:• Cellware stable for at least three
months at -20°C. Keep frozen until use.
• Thin layer cellware is stable for at least three months from date of shipment when stored at 2°C to 8°C.
Visit bdbiosciences .com/cellculturesystems
for more information on BD BioCoat matrigel
matrix cellware.
Multiwell Plates
description qty. cat.no.
6-well 2 354432
12-well 2 354503
24-well 2 354433
48-well 2 354508
Culture Dishes
description qty. cat.no.
35 mm 8 354460
Thin-Layer Multiwell and Assay Plates
description qty. cat.no.
6-well 5 354603
24-well 5 354605
96-well 5 354607
Thin-Layer Culture Dishes
description qty. cat.no.
35 mm 20 354602
60 mm 20 354601
100 mm 10 354600
For Hepatocytes
description qty. cat.no.
6-well plates 5 354510
100 mm culture dishes 5 354634
ES Culture Plates
description qty. cat.no.
6-well 5 354671
Rat Hepatocytes Cultured in the BD BioCoat Hepatocyte Differentiation Environment
Transmission electron micrograph of thin sections shows similar intracellular and intercellular
structures, indicative of healthy differentiated hepatocytes, in four-week-old cultures.
Intracellular structures active nucleus (5000x) Numerous mitochondria with calcium deposits; rough ER stacks near cell surfaces and surrounding mitochondria; glycogen stores; golgi, ribosomal rosette, lipid droplets.
Intercellular structures (8600x) Frequent interdigitation of apposing cells; gap and tight junctions; intercellular lumens with microvilli, characteristic of bile canaliculi.
BD BioCoat Matrigel Matrix Cellware
bdbiosciences.com20 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Cell Culture (continued)
bdbiosciences.com21Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Laminin (LM), a major component of base-
ment membranes, is a multifunctional
glycoprotein that is used as a substrate to
culture and maintain differentiated func-
tion of a wide variety of cells. Laminin
has been shown in culture to stimulate
neurite outgrowth, promote cell attach-
ment, chemotaxis, cell differentiation, and
neuronal survival.
BD BioCoat Laminin cellware applications include:
• Promotion of cell attachment and spreading
• Induction of cell differentiation and neurite outgrowth
• Increase of proliferation of myoblasts32
• Study of effects of laminin on cell behavior
• Cell adhesion assays
BD BioCoat Laminin cellware has been used to culture:
• SH-SY5Y (human neuroblastoma), Neuro-2A (mouse neuroblastoma), N1-E115 (rat neuroblastoma)33
• MCF-10A cells34,35
• SK-MEL-28 cells16
• HVSMC36
• MDA-231 breast cancer cell line21
Source:• EHS mouse tumor
Quality Control:• Tested for ability to initiate
differentiation (neurite outgrowth) of NG-108 rat glioma/mouse neuroblastoma cells
• Tested and found negative for bacteria and fungi
• Laminin purity >90% by SDS-PAGE (contains entactin)
Storage and Stability:• Stable for at least three months from
date of shipment when stored at 2°C to 8°C. Do not freeze.
Multiwell and Assay Plates
description qty. cat.no.
6-well 5 354404
12-well 5 354502
24-well 5 354412
48-well 5 354507
96-well 5 354410
Culture Dishes
description qty. cat.no.
35 mm 20 354458
60 mm 20 354405
100 mm 10 354452
150 mm 5 354553
Flasks
description qty. cat.no.
25 cm2 plug-seal† 10 354533
75 cm2 plug-seal†† 10 354522
† BD BioCoat 25 cm2 flasks are 70 mL canted neck
†† BD BioCoat 75 cm2 flasks are 250 mL canted neck
Effects of BD BioCoat Laminin Cellware on NG-108 Rat Glioma/Mouse Neuroblastoma Cells
NG-108 rat glioma/mouse neuroblastoma cells cultured on tissue culture plastic are loosely adhered and remain rounded.
NG-108 rat glioma/mouse neuroblastoma cells cultured on BD BioCoat Laminin cellware exhibit a spindle-shaped morphology and dendritic processes.
BD BioCoat Laminin Cellware
Type IV collagen is a ubiquitous component
in basement membranes and provides the
major structural support for this matrix.
When the collagen IV meshwork is assem-
bled, it provides a scaffold for the assembly
of other basement membrane components
through interactions with laminin, entactin/
nidogen, and heparan sulfate proteoglycan.
Collagen IV is useful as a substrate for
growth of nerve, epithelial, endothelial, and
muscle cells. Collagen plays a role in the
regulation of cell growth, differentiation
and adhesion, and tissue formation.
BD BioCoat™ Collagen IV cellware applications include:
• Promotion of cell attachment and spreading
• Cell differentiation and neurite outgrowth
• Increased proliferation of PC12 cells
• Studies of effects of collagen IV on cell behavior
• Cell adhesion assays
BD BioCoat Collagen IV cellware has been used to culture:
• PC12 cells14,37,38
• SH-SY5Y cells14
• Human melanoma cells lines SK-MEL-28-N1 and SK-MEL-2816
• Primary murine hepatocytes39
Source:• EHS lathrytic mouse tumor
Quality Control:• Tested for ability to initiate
differentiation (neurite outgrowth) of NG-108 rat glioma/mouse neuroblastoma cells
• Tested and found negative for bacteria and fungi
• Collagen IV purity >90% by SDS-PAGE
Storage and Stability:• Stable for at least three months from
date of shipment when stored at 2°C to 8°C.
bdbiosciences.com22 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Multiwell and Assay Plates
description qty. cat.no.
6-well 5 354428
24-well 5 354430
96-well 5 354429
Culture Dishes
description qty. cat.no.
35 mm 20 354459
60 mm 20 354416
100 mm 10 354453
150 mm 5 354554
Flasks
description qty. cat.no.
25 cm2 plug-seal† 10 354534
75 cm2 plug-seal†† 10 354523
175 cm2 plug-seal 5 354528
† BD BioCoat 25 cm2 flasks are 70 mL canted neck
†† BD BioCoat 75 cm2 flasks are 250 mL canted neck
Effects of BD BioCoat Collagen IV Cellware on PC12 Rat Pheochromocytoma Cells
PC12 cells cultured on tissue culture plastic do not attach well and tend to float in clumps in the culture medium.
PC12 cells cultured on BD BioCoat Collagen IV cellware show 90% attachment and rapid proliferation.
BD BioCoat Collagen IV Cellware
Cell Culture (continued)
bdbiosciences.com23Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Poly-D-Lysine (PDL) and Poly-L-Lysine (PLL)
are synthetic compounds that enhance cell
adhesion and protein absorption by altering
surface charges on the culture substrate. In
addition to promoting cell adhesion, poly-
lysine surface treatments support neurite
outgrowth and improve the survival of
many central nervous system (CNS) primary
cells in culture. Because PDL and PLL are
synthetic molecules, they do not stimulate
biological activities that occur in vivo, and
they do not introduce impurities carried by
natural polymers.
BD BioCoat Poly-Lysine cellware applications include:
• Attachment and spreading of a variety of cell types
• Cell differentiation and neurite outgrowth
• Attachment of fastidious transfected cell lines
• Support survival of primary neurons in culture
• Serum-free or reduced serum culture
BD BioCoat Poly-Lysine cellware has been used to culture:
• Primary mouse brain capillaries40
• HEK-293 cells41-44
• MDA-231 breast cancer cells21
• Mouse cerebellar granule neurons45
• Transfected rat 1 cells46
• Rat anterior pituitary cells47
• Transfected COS-7 cells48
• Transiently transfected primary rat astrocytes49
• Rat primary cerebellar granule neurons50,51
• Murine microglia MG-7 cells52
Source:• PDL, synthetic (MW 75–150 kD)
• PLL, synthetic (MW 30–70 kD)
Storage and Stability:• Cellware stable for at least six months
from date of shipment when stored at 4°C to 30°C under dry conditions.
• Coverslips and culture slides stable for at least three months from date of shipment when stored at 2°C to 8°C.
Quality Control:• Multiwell plates (6, 12, 24, 48-well),
dishes, flasks, coverslips, and culture slides tested for ability to promote firm attachment of RCG cells.
• Assay plates (96- and 384-well) tested for signal from EcoPack™ 2-293 cells cultured in serum-free medium for one day, washed on a Skatron washer (Molecular Devices), and labeled with Calcein-AM. Intra- and inter-plate percent CVs are measured to ensure consistent coating. EcoPack 2-293 is a transformed HEK-293 cell line.
• All lots tested and found negative for bacteria and fungi.
Effect of BD BioCoat PDL on Cortical Neurons
Effect of BD BioCoat PDL on Rat Cerebellar Granule (RCG) Cells
Mixed culture of cortical neurons and astro-cytes cultured on BD BioCoat PDL cellware. Neurons are highly branched with very long processes. Astrocytes show similar process elongation.
RCG cells cultured on BD BioCoat PDL cell-ware show firm attachment (similar results obtained on PLL).
BD BioCoat Poly-Lysine Cellware
BD BioCoat Poly-D-Lysine Cellware Multiwell and Assay Plates
description qty. cat.no. qty. cat.no. qty. cat.no.
6-well 5 354413 50 356413
12-well 5 354470 50 356470
24-well 5 354414 50 356414
48-well 5 354509 50 356509
96-well Clear 5 354461 50 356461 80 356690
96-well Black/Clear 5 354640 50 356640 80 356692
96-well White/Clear 5 354651 50 356651 80 356693
96-well White 5 354620 50 356620 80 356691
384-well Black/Clear 5 354663 50 356663 80 356697
384-well White/Clear 5 354660 50 356660 80 356694
384-well Clear 5 354662 50 356662 80 356696
384-well White 5 354661 50 356661 80 356695
Coverslips (No . 1 German Glass)
description qty. cat.no.
12 mm Round 80 354086
Culture Dishes
description qty. cat.no. qty. cat.no.
35 mm 20 354467 100 356467
60 mm 20 354468 100 356468
100 mm 10 354469 40 356469
150 mm 5 354550
Coverslip-Bottom Dishes (No . 1 German Glass)
description qty. cat.no.
35 mm dish 20 354077with coverslip bottom
Flasks
description qty. cat.no. qty. cat.no.
25 cm2 plug-seal† 10 354479 50 356479
25 cm2 vented-cap† 10 354536 50 356536
75 cm2 plug-seal†† 5 354524 50 356524
75 cm2 vented-cap†† 5 354537 50 356537
150 cm2 plug-seal 5 354495 40 356495
150 cm2 vented-cap 5 354538 40 356538
175 cm2 plug-seal 5 354529 40 356529
175 cm2 vented-cap 5 354539 40 356539
† BD BioCoat 25 cm2 flasks are 70 mL canted neck
†† BD BioCoat 75 cm2 flasks are 250 mL canted neck
CultureSlides
description qty. cat.no.
1-well 12 354566
2-well 12 354629
4-well 12 354577
8-well 12 354632
BD BioCoat Poly-L-Lysine Cellware Multiwell and Assay Plates
description qty. cat.no. qty. cat.no.
6-well 5 354515 50 356515
96-well Clear 5 354516 50 356516
Culture Dishes
description qty. cat.no. qty. cat.no.
35 mm 20 354518 100 356518
60 mm 20 354517 100 356517
Coverslips (No . 1 German Glass)
description qty. cat.no.
12 mm Round 80 354085
bdbiosciences.com24 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
BD BioCoat Poly-Lysine Cellware (continued)
Cell Culture (continued)
bdbiosciences.com25Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
For some applications, the use of a combi-
nation of ECM proteins, such as laminin
(LM) and human fibronectin (HFN), or LM
and attachment factors such as Poly-D-
Lysine (PDL) or Poly-L-Orinthine (PLO) has
been shown superior to the use of either
alone.
BD BioCoat™ PDL/LM and PLO/LM cellware
is suitable for culturing many different types
of peripheral nervous system (PNS) and
central nervous system (CNS) networks,
and is useful for promoting neural cell
attachment and differentiation. BD BioCoat
LM/HFN cellware provides an in vitro envi-
ronment that promotes cell attachment and
extensive process formation.
BD BioCoat PDL/LM, PLO/LM, and LM/HFN cellware applications include:
• Enhancement of neuronal cell attachment to plastic and glass
• Promotion of neurite outgrowth
• Culture of glial cells as a feeder layer for neurons
• Construction of neuronal cell model systems to study CNS function, development, and diseases
BD BioCoat PDL/LM has been used to culture:
• SH-SY5Y (human neuroblastoma), Neuro-2A (mouse neuroblastoma), N1-E115 (rat neuroblastoma)53
• Primary rat hippocampus54
• Murine T11-L3 dorsal root ganglion neurons (DRGNs)55,56
• Transfected PC12 cells56
• MCF-10A cells35
• Rat primary DRGNs57
Source:• PDL, synthetic (MW 75-150 kD)
• PLO, synthetic (MW 30-70 kD)
• LM, EHS mouse tumor
• HFN, human plasma
NOTE: Source material tested for hepatitis B antigen and HIV-1 antibody
Storage and Stability:• Stable for at least three months from
date of shipment when stored at 2°C to 8°C. Do not freeze.
Quality Control:• PDL/LM and PLO/LM tested for ability
to initiate differentiation (neurite outgrowth) of NG-108 rat glioma/mouse neuroblastoma cells
• Tested and found negative for bacteria and fungi
• LM/HFN tested for receptor agonist induced changes in intracellular calcium using Fluo-3A (a long wavelength fluorescent calcium indicator) in primary rat cortical neuron enriched cultures58,59
Multiwell and Assay PlatesPDL/LM
description qty. cat.no.
6-well 5 354595
24-well 5 354619
96-well 5 354596
Multiwell and Assay Plates (continued) PLO/LM
description qty. cat.no.
6-well 5 354658
24-well 5 354659
96-well 5 354657
LM/HFN
description qty. cat.no.
96-well 5 354670
Culture DishesPDL/LM
description qty. cat.no.
100 mm 10 354455
Coverslips (No . 1 German Glass)PDL/LM
description qty. cat.no.
12 mm round 80 354087
CultureSlidesPDL/LM
description qty. cat.no.
2-well 12 354687
8-well 12 354688
Effects of BD BioCoat LM/HFN on Primary Neurons
Neurotransmitter induced calcium released in rat primary neurons plated on BD BioCoat LM/HFN Cellware.
BD BioCoat Poly-D-Lysine/Laminin, Poly-L-Orinthine/Laminin, and Laminin/Human Fibronectin Cellware
Collagen I, found in most tissues and
organs, is most plentiful in dermis, tendon,
and bone. It is an integral part of the frame-
work that holds cells and tissues together
and has been recognized as a useful matrix
for improving cell culture. In vitro use of
collagen can exert effects on the adherence,
morphology, growth, migration, and differ-
entiation of a variety of cell types.11
BD BioCoat Collagen I cellware applications include:
• Promotion of cell attachment and spreading
• Rapid expansion of cell populations
• Serum-free or reduced serum culture
• Cell adhesion assays improving survival of primary cells in culture
BD BioCoat Collagen I cellware has been used to culture:
• Primary murine cardiac myocytes12
• Human vascular SMC13
• PC12 cells and SH-SY5Y cells14
• Mouse primary keratinocytes15
• SK-MEL-28-N1 cells16
• Murine myoblast C2C12 cells17
• HUVEC18
• HEK-293 cells19
• Rat Kupffer cells20
• MDA-231 breast cancer cells21
Source:• Rat tail tendon
Quality Control:• Tested for ability to promote
attachment and spreading of HT-1080 human fibrosarcoma cells
• Tested and found negative for bacteria and fungi
• Collagen I purity >90% by SDS-PAGE
Storage and Stability:• Cellware stable for at least six months
from date of shipment when stored at 4°C to 30°C under dry conditions.
• Coverslips and CultureSlides stable for at least three months from date of shipment when stored at 2°C to 8°C.
CultureSlides
description qty. cat.no.
1-well 12 354556
2-well 12 354627
4-well 12 354557
8-well 12 354630
Coverslips (No . 1 German Glass)
description qty. cat.no.
22 mm Round 60 354089
Flasks
description qty. cat.no. qty. cat.no.
25 cm2 plug-seal 10 354531 50 356531
25 cm2 vented-cap 10 354484 50 356484
75 cm2 plug-seal 5 354462 50 356462
75 cm2 vented-cap 5 354485 50 356485
150 cm2 plug-seal 5 354645 40 356645
150 cm2 vented-cap 5 354486 40 356486
175 cm2 plug-seal 5 354478 40 356478
175 cm2 vented-cap 5 354487 40 356487
Culture Dishes
description qty. cat.no. qty. cat.no.
35 mm 20 354456 100 356456
60 mm 20 354401 100 356401
100 mm 10 354450 40 356450
150 mm 5 354551
Multiwell and Assay Plates
description qty. cat.no. qty. cat.no. qty. cat.no.
6-well 5 354400 50 356400
12-well 5 354500 50 356500
24-well 5 354408 50 356408
48-well 5 354505 50 356505
96-well Clear 5 354407 50 356407 80 356698
96-well Black/Clear 5 354649 50 356649 80 356700
96-well White/Clear 5 354650 50 356650 80 356701
96-well White 5 354519 50 356519 80 356699
384-well Black/Clear 5 354667 50 356667 80 356705
384-well White/Clear 5 354664 50 356664 80 356702
384-well Clear 5 354666 50 356666 80 356704
384-well White 5 354665 50 356665 80 356703
Please call for shipping on large orders
Effects of BD BioCoat Collagen I Cellware on Fetal Bovine Heart Endothelial (FBHE) Cells
FBHE cells grown for five days in basal medium containing 10% FBS on tissue culture plastic show sparse growth.
FBHE cells grown for five days using the BD BioCoat Endothelial Cell Growth Environment (Collagen I Cellware) form a confluent monolayer and show numerous mitotic cells.
BD BioCoat Collagen I Cellware
bdbiosciences.com26 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Cell Culture (continued)
Human fibronectin (HFN) is a widely distrib-
uted glycoprotein that is used as a substrate
to promote attachment of cells through its
integrin-binding arginine-glycine-aspartate
(RGD) motif. HFN is a product of most
mesenchymal and epithelial cells and is
present in both the extracellular matrix
(ECM) and plasma. The principal function
of HFN appears to be in cellular migration
during wound healing and development,
regulation of cell growth and differentia-
tion, and haemostasis/thrombosis.
BD BioCoat Fibronectin cellware applications include:
• Promotion of cell attachment and spreading
• Rapid expansion of cell populations
• Serum-free or reduced serum culture
• Cell adhesion assays
• Study of effects of HFN on cell behavior
• Improve survival of primary cells in culture
BD BioCoat Fibronectin cellware has been used to culture:
• 3T3 Preadipocytes60
• Transfected 293T and transfected H1299 cells61
• MCF-10A cells34
• Primary cord blood mononuclear cells62
• SK-MEL-28 (human melanoma cells)16
• NIH3T3 cells63
• MDA-231 human breast cancer cells21
Source:• Human plasma
NOTE: Source material tested for hepatitis B antigen and HIV-1 antibody
Quality Control:• Tested for ability to promote
attachment and spreading of BHK-21 hamster kidney cells
• Tested and found negative for bacteria and fungi
• Fibronectin purity >90% by SDS-PAGE
Storage and Stability:• Stable for at least three months from
date of shipment when stored at 2°C to 8°C. Do not freeze.
Multiwell and Assay Plates
description qty. cat.no.
6-well 5 354402
12-well 5 354501
24-well 5 354411
48-well 5 354506
96-well 5 354409
† BD BioCoat 25 cm2 flasks are 70 mL canted neck
†† BD BioCoat 75 cm2 flasks are 250 mL canted neck
Culture Dishes
description qty. cat.no.
35 mm 20 354457
60 mm 20 354403
100 mm 10 354451
150 mm 5 354552
Flasks
description qty. cat.no.
25 cm2 plug-seal† 10 354532
75 cm2 plug-seal†† 10 354521
150 cm2 plug-seal 5 354646
175 cm2 plug-seal 5 354526
Coverslips (No . 1 German Glass)
description qty. cat.no.
22 mm round 60 354088
CultureSlides
description qty. cat.no.
1-well 12 354558
2-well 12 354628
4-well 12 354559
8-well 12 354631
Effects of BD BioCoat Fibronectin Cellware on BHK-21 Cells
BHK-21 fibroblasts cultured on glass CultureSlides do not spread.
BHK-21 fibroblasts cultured on BD BioCoat HFN CultureSlides attach and spread within one hour.
BD BioCoat Fibronectin Cellware
bdbiosciences.com27Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
bdbiosciences.com28 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
BD BioCoat™ Gelatin cellware provides
an attachment and growth-promoting
substrate for the culture of a variety of
cell types. Gelatin is used commonly in
the culture of vascular endothelial cells,
muscle, embryonic stem (ES) cells, and F9
teratocarcinoma cells. It is also suitable
for promoting adhesion of transfected cell
types. Gelatin is a heterogeneous mixture of
water-soluble proteins derived through the
hydrolysis of collagen.
BD BioCoat Gelatin cellware applications include:
• Promotion of cell attachment and spreading
• Culture of normal and transfected F9 teratocarcinoma cells for gene expression studies65
• Culture of human umbilical vein endothelial cells (HUVECs) for E-Selectin66 expression and VEGF induction
BD BioCoat Gelatin cellware has been used to culture:
• HUVEC,66 BME,22 BAEC23
• ES cells24
• C2C1235 and MM1464 myoblasts
• Normal and transfected F9 teratocarcinoma cells
Source:• Gelatin, porcine
Quality Control:• Tested for ability to promote
proliferation of HUVECs
• Tested and found negative for bacteria and fungi
Storage and Stability:• Stable for at least six months from date
of shipment when stored at 4°C to 30°C under dry conditions.
Multiwell and Assay Plates
description qty. cat.no. qty. cat.no.
6-well 5 354652 50 356652
96-well 5 354689 50 356689
Culture Dishes
description qty. cat.no. qty. cat.no.
100 mm 10 354653 40 356653
Culture Flasks
description qty. cat.no. qty. cat.no.
75 cm2 plug-seal† 5 354654 50 356654
75 cm2 vented cap† 5 354488 50 356488
†BD BioCoat 75 cm2 flasks are 250 mL canted neck
Effect of BD BioCoat Gelatin Cellware on Human Umbilical Vein Endothelial Cells (HUVECs)
HUVECs grown for seven days on BD BioCoat Gelatin 6-well multiwell plates seeded at a density of 2×104 in the presence of BD™ Endothelial Cell Culture Medium.
BD BioCoat Gelatin Cellware
Cell Culture (continued)
bdbiosciences.com29Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
BD BioCoat Custom Coating Service
Custom coating, the way you want it.
BD BioCoat Custom Coating Service offers
an extensive selection of cell-based coatings
on a wide variety of BD Falcon vessels—
from roller bottles to flasks to 384-well
plates. We can even develop special coat-
ings for you. Simply tell us the type of
vessel and coating you would like applied
and we’ll make a trial sample for you to
evaluate.
BD BioCoat Consulting Service
BD Biosciences is committed to helping you
optimize your cell culture conditions. The
process of selecting the most appropriate
substratum is often empirical and time
consuming. Our consulting service can
facilitate this process for you. Our highly
trained technical support staff will assist you
in determining which BD BioCoat product is
best for your cell type and application. Let
BD Biosciences put its comprehensive tech-
nical database and expertise in cell culture
to work for you.
BD BioCoat Bar-Coding Service
Get on the road to increased productivity
with barcoded BD BioCoat cellware. The
BD Bar-Coding Service provides high-quality
bar code labels affixed to any side of a
microplate. Bar codes have been quality
tested for optimal readability, chemical resis-
tance, and temperature durability.
BD Falcon and BD BioCoat cell culture inserts and insert systems
combine the microporous polyethylene terephthalate (PET)
membrane with multiple formats and pore sizes for a variety of
cell-based applications. A number of insert model systems have
been developed to study cells that are derived from the brain and
peripheral nervous system. For example, the BD Falcon FluoroBlok™
24-Multiwell Insert System has been used to examine dorsal root
ganglion neurite formation in an automated, fluorescence-based
assay.67 In addition to this application, BD cell culture insert products
are available for establishing co-culture systems and other models to
study the functional properties of neuronal and glial cells.
BD Falcon Cell Culture Inserts0.4 µm pore size
description qty./pack qty./case cat.no.
inserts for 6-well plates 1 48 353090
inserts for 12-well plates 1 48 353180
inserts for 24-well plates 1 48 353095
BD Falcon FluoroBlok Cell Culture Inserts
description qty./pack qty./case cat.no.
1.0 µm inserts for 24-well plates 1 48 351150
3.0 µm inserts for 24-well plates 1 48 351151
8.0 µm inserts for 24-well plates 1 48 351152
BD Falcon FluoroBlok 24-Multiwell Insert Systemswith Feeder Tray and Lid
description qty./pack cat.no.
1.0 µm pore size 1 351153
5 351154
BD Falcon FluoroBlok 24-Multiwell Insert Systemswith 24-well plate and lid
description qty./pack cat.no.
3.0 µm pore size 1 351155
5 351156
8.0 µm pore size 1 351157
5 351158
BD Falcon FluoroBlok 96-Multiwell Insert Systems
description qty./pack cat.no.
3.0 µm pore size 1 351161
5 351162
8.0 µm pore size 1 351163
5 351164
Visit bdbiosciences .com/cellcultureinserts for additional information about our cell culture inserts and insert systems.
BD BioCoatSM Services
Cell Culture Inserts and Insert Systems
Visit bdbiosciences .com/custom for additional information about our BD BioCoatSM Services.
bdbiosciences.com30 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Growth FactorsEpidermal Growth Factors
description qty. cat.no.
BD EGF, mouse natural (culture grade) 100µg 354001
(10x100µg) 100µg 356001
BD EGF, mouse natural (receptor grade) 100µg 354010
(5x100µg) 500µg 356010
BD EGF, human recombinant 100µg 354052
(10x100µg) 100µg 356052
Fibroblast Growth Factors
description qty. cat.no.
BD bFGF, bovine natural 10µg 356037
BD bFGF, human recombinant 10µg 354060
(5x10µg) 50µg 356060
(10x10µg) 100µg 356061
Insulin-like Growth Factor
BD IGF-I, human recombinant (culture) 10µg 354037
Nerve Growth Factors
description qty. cat.no.
BD 2.5S NGF, mouse natural 10µg 354005
100µg 356004
(2x500µg) 100µg 356005
BD 7S NGF, mouse natural 100µg 354009
Platelet-Derived Growth Factor
description qty. cat.no.
BD PDGF-BB, human recombinant 10µg 354051
(10x10µg) 100µg 356051
Vascular Endothelial Growth Factor
description qty. cat.no.
BD VEGF, human recombinant 10µg 354107
Transforming Growth Factor
description qty. cat.no.
BD TGF-b, human natural 1µg 354039
(5x1µg) 5µg 356039
(5x2µg) 10µg 356040
From purified growth factors to enriched culture supplements and
speciality media, these products all allow you to propagate your cells
under more defined serum-reduced or serum-free conditions.
BD Biosciences supplies a comprehensive line of high-quality cyto-
kines and media additives to meet your individual needs for culturing
animal or human cells in vitro:
• Vialed, highly purified growth factors with high biological activity
• Enriched culture supplements to be reconstituted in the medium of your choice
Lymphokines• Broad range of highly purified natural and
recombinant lymphokines
• Membrane filtered (0.2 μm membrane) and tested for biological activity
• High specific activity
• Available as vialed pure factors or as enriched conditioned media
Media Additives• Serum-free or low-serum alternatives to routinely used
high serum containing media
• Supplements and essential components used as the basis of specialized serum-free media
• Membrane filtered (0.2 μm membrane) and tested for biological activity
BD™ Growth Factors
Cell Culture (continued)
description qty. cat.no.
LymphokinesInterleukin-1
description qty. cat.no.
BD IL-1b, human recombinant 2µg 354042
Interleukin-2
description qty. cat.no.
BD IL-2, human recombinant BRMP*units 10,000 354043
BRMP*units 50,000 356043
BD IL-2, rat natural BRMP*units 4,000 354110
BD IL-2, mouse recombinant BRMP*units 10,000 356078
BRMP*units 25,000 354078
Interleukin-3
description qty. cat.no.
BD IL-3, mouse recombinant 10µg 354058
Interleukin-4
description qty. cat.no.
BD IL-4, human recombinant 5µg 354068
Granulocyte-Macrophage Colony Stimulating Factor
description qty. cat.no.
BD GM/CSF, human recombinant 1µg 354048
Tumor Necrosis Factor-a
description qty. cat.no.
BD TNF-a, human recombinant 10µg 354066
(5x10µg) 50µg 356066
Stem Cell Factor
description qty. cat.no.
BD SCF, human recombinant 10µg 354105
Media Additives and SupplementsBD Nu-Serum Serum Replacements
description qty. cat.no.
BD Nu-Serum™ Serum Replacements 100mL 355100
500mL 355500
BD Nu-Serum™ IV Serum Replacements 100mL 355104
500mL 355504
BD Nu-Serum™ V Serum Replacements 100mL 355105
500mL 355505
Media Additives and Supplements (continued) T-Cell Culture Supplements
description qty. cat.no.
BD T-Cell Culture Supplement with ConA, rat (IL-2 culture supplement)
100mL 354115
BD T-Cell Culture Supplement without ConA, rat (IL-2 culture supplement)
100mL 354116
IL-3 Culture Supplement
description qty. cat.no.
BD IL-3 Culture Supplement, mouse 25mL 354040
Other Media Additives
description qty. cat.no.
BD Albumin, bovine serum (BSA, delipidized) 10g 354331
BD Linoleic Acid/Albumin Complex Linoleicacid/BSA 2.5/500mg 354227
BD Hydrocortisone 50mg 354203
BD Selenous Acid (sodium salt) 100mg 354201
BD Transferrin, human (holo) 100mg 354204
1g 354304
ITS Universal Culture Supplements
description qty. cat.no.
BD ITS Premix (5literequivalent) 5mL 354351
(20literequivalent) 20mL 354350
BD ITS+ Premix (2literequivalent) 20mL 354352
Mito+ Serum Extender
description qty. cat.no.
BD Mito+ Serum Extender (5literequivalent) 5mL 355006
Bovine Pituitary Extract
description qty. cat.no.
BD BPE 1.5mg 354123
(5x1.5mg) 7.5mg 356123
description qty. cat.no.
Endothelial Cell Growth Supplement
BD ECGS 1.5mg 354006
100mg 356006
bdbiosciences.com31Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
* benzamide ribose monophosphate
bdbiosciences.com32 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Functional Assays and Discovery Reagents
Programmed cell death, or apoptosis, is a normal physiological
process that occurs during embryonic development and in the
maintenance of tissue homeostasis. The term apoptosis, from the
Greek word for the “falling off” of leaves from a tree, is used to
describe a process in which a cell actively participates in its own
destructive processes. The apoptotic program is characterized
by certain morphological features. These include changes in the
plasma membrane such as loss of membrane symmetry and loss
of membrane attachment, a condensation of the cytoplasm and
nucleus, and internucleosomal cleavage of DNA. In the final stages,
the dying cells become fragmented into “apoptotic bodies,”
which are rapidly eliminated by phagocytic cells without eliciting
significant inflammatory damage to surrounding cells. Inappropriate
induction of apoptosis has widespread pathological implications
and has been associated with diseases such as neurodegenerative
diseases, graft-versus-host diseases, transplant rejection, cancer,
and AIDS.73-77 The study of apoptosis has never been more important
than it is today. Along with this intensive study has come the
development of a multitude of methods to detect apoptotic cells
and dissect the mechanisms of apoptosis. It is important to note that
not every cell type will display all the classical features of apoptosis.
As such, studies designed to measure multiple aspects of apoptosis
may be required to analyze and confirm the mechanism of cell death.
BD Biosciences offers a full line of apoptosis detection research tools
and technologies for measuring indicators of apoptosis at different
stages. These reagents and technologies are based on the following
distinctive features of apoptosis:
• Plasma membrane alterations
• Mitochondrial changes
• Activation of caspases
• DNA fragmentation
For a complete list of over 400 apoptosis-related research reagents
and kits, visit bdbiosciences .com/literature to request our “Life,
Death, and Cell Proliferation” brochure.
Apoptosis
bdbiosciences.com33Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Cell Signaling*
description react clone isotype apps format size cat.no.
14-3-3 Hu C23-1 MouseIgG1 WB Purified 100µg 556596
14-3-3epsilon Chick,Dog,Frog,Hu,Ms,Rat 12 MouseIgM IP,IF,IHC,WB Purified 50µg 610542
IP,IF,IHC,WB Purified 150µg 610543
Akt (PKBa) Dog,Hu,Ms,Rat 2 MouseIgG2a IF,WB Purified 150µg 610877
7 MouseIgG1 IF,WB Purified 150µg 610837
55 MouseIgG1 IF,WB Purified 150µg 610861
Calreticulin Hu,Ms,Rat 16 MouseIgG1 IF,WB Purified 150µg 612137
CAPS Ms,Rat 4 MouseIgG1 WB Purified 50µg 612138
Caveolin (pY14) Hu,Ms,Rat 56 MouseIgG1 IF,IHC,WB Purified 150µg 611339
Caveolin 1 Chick,Hu 2234 MouseIgG2a IF,IP,WB Purified 150µg 610494
Chick,Dog,Hu,Ms,Rat 2297 MouseIgG1 FCM,IF,IHC,IP,WB Purified 150µg 610407
Dog,Hu,Ms,Rab,Rat C060 MouseIgM FCM,IF,IHC,IP,WB Purified 150µg 610058
Chick C20B MouseIgG1 IF,IP,WB Purified 150µg 610388
Caveolin 3 Ms,Rab,Rat 26 MouseIgG1 IHC,WB Purified 150µg 610421
cGB-PDE Rat 54 MouseIgG1 IF,WB Purified 50µg 611498
IF,WB Purified 150µg 611499
Dlg Hu,Ms,Rat 12 MouseIgG1 IF,IP,WB Purified 50µg 610874
IF,IP,WB Purified 150µg 610875
Doublecortin Ms 30 MouseIgG1 IF,WB Purified 50µg 611706
Gat Chick,Hu,Ms,Rat 3 MouseIgG1 IF,IHC,WB Purified 150µg 610589
GAP-43/Neuromodulin Hu,Ms,Rat 31 MouseIgG1 WB Purified 50µg 612262
WB Purified 150µg 612263
JBP1 (Skb1Hs) Dog,Hu,Ms,Rat 32 MouseIgG1 IF,WB Purified 150µg 611539
JIP-1 Ms,Rat 50 MouseIgG1 WB Purified 150µg 611891
Jun Bov,Chick,Dog,Hu,Ms,Rat 3 MouseIgG2a IF,IHC,IP,WB Purified 150µg 610327
N-Copine Ms,Rat 42 MouseIgG1 IF,WB Purified 50µg 611588
Dros 42 MouseIgG1 IF,WB Purified 50µg 612638
NCS-1 Hu,Ms,Rat 1 MouseIgG1 IF,WB Purified 50µg 611578
Basic classifications of protein kinases fall into these key areas of
research:
• G-protein coupled receptors
• G-protein regulators
• Intracellular hormone receptors
• Receptor tyrosine kinases
• Non-receptor tyrosine kinases
• Receptor serine/threonine kinases
• Non-receptor serine/threonine kinases
• Phospholipids and phospholipases
• Phosphatases
Signal transduction at the cellular level is the transference of external
cell signals to internal mechanisms. Simple signal transduction uses
receptor molecules such as the acetylcholine class: receptors that
constitute channels which allow signals to be passed in the form of
small ion movement, either into or out of the cell (see Ion Channels
and Transporter on page 50). Complex signal transduction relies on
the coupling of ligand-receptor interactions. Resulting events include
phosphorylations by tyrosine kinases and/or serine/threonine kinases.
Protein phosphorylations change enzyme activities and protein
conformations. The resulting alteration in cellular activity changes
the programming of gene expression and the up- and down-regu-
lation of mechanistic proteins. One of the key receptor classes in
neurobiology is tyrosine kinases (RTKs)—neurotrophin receptor
family (trkA, trkB, trkC) and NGF receptor.
Cell Signaling, Including Kinases and Phosphotases
*Visit bdbiosciences .com for a complete listing of cell signaling reagents.
Cell Signaling* (continued)
description react clone isotype apps format size cat.no.
nNOS/NOS Type I Dog,Hu,Ms,Rat 16 MouseIgG2a IF,IHC,IP,WB Purified 50µg 610308
IF,IHC,IP,WB Purified 150µg 610309
Rat 52 MouseIgG1 WB Purified 50µg 611852
WB Purified 150µg 611853
Hu,Ms,Rat Polyclonal RabbitIg IF,IHC,IP,WB Purified 50µg 610310
IF,IHC,IP,WB Purified 150µg 610311
p116 Rip Dog,Hu,Ms,Rat 43 MouseIgG1 IF,WB Purified 50µg 611098
p38a (SAPK2a) Dog,Hu,Ms,Rat 27 MouseIgG1 IF,WB Purified 150µg 612169
p62dok Hu 45 MouseIgG1 WB Purified 150µg 610753
p96 (Disabled-2) Hu,Ms,Rat 52 MouseIgG1 IF,WB Purified 150µg 610465
Phocein Rat 38 MouseIgG2b IF,WB Purified 50µg 612694
IF,WB Purified 150µg 612695
PIN Hu,Rat 4 MouseIgG1 IF,IP,WB Purified 50µg 610726
IF,IP,WB Purified 150µg 610727
PITPalpha Dog,Hu,Ms,Rab,Rat 50 MouseIgG1 IF,WB Purified 50µg 611024
PITPNM Ms,Rat 8 MouseIgG1 WB Purified 50µg 612026
WB Purified 150µg 612027
RACK1 Bov,Chick,Dog,Frog,Hu,Ms,Rat 20 MouseIgM IF,IHC,IP,WB Purified 150µg 610178
Ras Chick,Dog,Hu,Ms,Rat 18 MouseIgG1 IF,IHC,IP,WB Purified 150µg 610002
Ras-GRF2 Hu,Ms,Rat 23 MouseIgG1 IF,WB Purified 150µg 610841
SH2-B Hu,Ms,Rat 46 MouseIgG1 IF,WB Purified 50µg 611526
IF,WB Purified 150µg 611527
ShcC Hu,Ms,Rat 23 MouseIgG1 IF,IP,WB Purified 50µg 610642
IF,IP,WB Purified 150µg 610643
SHPS-1 Rat 17 MouseIgG1 WB Purified 150µg 611497
SPA-1 Hu,Ms,Rat 3 MouseIgG1 IF,WB Purified 150µg 611215
Stat1 (C-Terminus) Chick,Dog,Hu,Ms,Rat 42 MouseIgG2b IF,IP,WB Purified 150µg 610186
Stat1 (N-Terminus) Chick,Dog,Frog,Hu,Ms,Rat 1 MouseIgG1 WB HRP 150µg 610118
IF,IHC,IP,WB Purified 150µg 610116
Polyclonal RabbitIg IF,IP,WB Purified 150µg 610120
Stat1 (pY701) Hu,Ms 14 MouseIgG1 IF,IP,WB Purified 150µg 612133
4a MouseIgG2a IF,IP,WB Purified 150µg 612233
Stat2 Chick,Hu 22 MouseIgG2a IP,WB Purified 150µg 610188
Stat5 (MGF) Dog,Hu,Ms,Rat 89 MouseIgG2b FCM,IF,IHC,WB Purified 150µg 610192
Stat5 (pY694) Hu Polyclonal RabbitIg IF,WB Purified 150µg 611819
47 MouseIgG1 WB Purified 150µg 611965
Stat5A Dog,Hu,Ms,Rat 51 MouseIgG1 IF,WB Purified 150µg 611835
Stat6 (pY641) Hu 18 MouseIgG2a WB Purified 150µg 611567
Polyclonal RabbitIg WB Purified 150µg 611821
Striatin Dog,Hu,Ms,Rat 6 MouseIgG2b IF,WB Purified 50µg 610838
YT521-B Rat 8 MouseIgG1 IF,WB Purified 50µg 612380
ZO-1 Hu 1 MouseIgG1 IF,WB Purified 50µg 610966
Circadian Regulator
description react clone isotype apps format size cat.no.
Per2 Ms,Rat 52 MouseIgG1 IF,WB Purified 50µg 611138
*Visit bdbiosciences .com for a complete listing of cell signaling reagents.
bdbiosciences.com34 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Functional Assays and Discovery Reagents (continued)
Kinases and Phosphatases
description react clone isotype apps format size cat.no.
Calcineurin Chick,Frog,Hu,Ms,Rat 29 MouseIgG2a IF,IHC,WB Purified 150µg 610260
CaM Kinase Kinase Dog,Hu,Rat 6 MouseIgG2a IF,IHC,IP,WB Purified 50µg 610544
IF,IHC,IP,WB Purified 150µg 610545
CaM Kinase II Dros 45 MouseIgG1 IF,IHC,WB Purified 50µg 612624
Dog,Hu,Ms,Rat 45 MouseIgG1 IF,IHC,WB Purified 50µg 611292
IF,IHC,WB Purified 150µg 611293
CaM Kinase IV Hu,Ms,Rat 26 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610275
IF,IHC,IP,WB Purified 150µg 610276
CaM Kinase VI Ms,Rat 5 MouseIgG1 IF,WB Purified 50µg 611736
IF,WB Purified 150µg 611737
CASK Dog,Frog,Hu,Ms,Rat 7 MouseIgG1 IF,IP,WB Purified 50µg 610782
IF,IP,WB Purified 150µg 610783
c-Raf-1 Chick,Dog,Hu,Ms,Rat 53 MouseIgG1 IF,IP,WB Purified 150µg 610152
CRIK Hu,Ms,Rat 6 MouseIgG1 IF,WB Purified 50µg 611376
IF,WB Purified 150µg 611377
DARPP-32 Ms,Rat 15 MouseIgG1 IF,WB Purified 50µg 611520
DGKi Hu,Ms,Rat 25 MouseIgG1 WB Purified 50µg 612150
Dyrk Hu,Rat 106 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610674
IF,IHC,IP,WB Purified 150µg 610675
ERK (pan) Chick,Dog,Frog,Hu,Ms,Rat 16 MouseIgG2a IF,IHC,IP,WB Purified 150µg 610124
ERK1 Chick,Dog,Frog,Hu,Ms,Rat MK12 MouseIgG1 IF,IHC,IP,WB Purified 150µg 610031
ERK2 Chick,Dog,Frog,Hu,Ms,Rat 33 MouseIgG2b IF,IHC,IP,WB Purified 150µg 610104
GSK-3b Chick,Dog,Hu,Ms,Rat 7 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610201
IF,IHC,IP,WB Purified 150µg 610202
Dros 7 MouseIgG1 IF,IP,WB Purified 50µg 612634
GSK-3b (pY216) Hu,Ms,Rat 13a MouseIgG1 WB Purified 50µg 612312
WB Purified 150µg 612313
Inhibitor 2 Hu 52 MouseIgG1 IF,WB Purified 150µg 610699
LIMK1 Hu,Ms,Rat 42 MouseIgG1 IF,WB Purified 150µg 611749
MRCKalpha Rat 41 MouseIgG1 IF,WB Purified 50µg 611584
IF,WB Purified 150µg 611585
MYPT1 Hu,Ms,Rat 20 MouseIgG1 WB Purified 150µg 612165
Ntk Chick,Dog,Frog,Hu,Ms,Rat 32 MouseIgG1 IF,IHC,WB Purified 150µg 610343
p38d (SAPK4) Hu 20 MouseIgG1 IF,WB Purified 150µg 610953
PDK1 (PKB Kinase) Dog,Hu,Ms,Rat 5 MouseIgG1 IF,WB Purified 150µg 611071
PKAC Dog,Hu,Ms,Rat 5B MouseIgG2b IF,WB Purified 150µg 610981
PKARI Chick,Dog,Frog,Hu,Ms,Rat 18 MouseIgG2b IF FITC 150µg 610168
IF,IP,WB Purified 150µg 610166
PKARIa Chick,Dog,Hu,Ms,Rat 20 MouseIgG1 WB Purified 150µg 610610
PKARIIa Dog,Hu,Ms,Rat 40 MouseIgG1 IF,WB Purified 150µg 612243
PKARIIb Chick,Dog,Hu,Ms,Rat 45 MouseIgG1 IF,IHC,IP,WB Purified 150µg 610626
PKCa Chick,Dog,Frog,Hu,Ms,Rat 3 MouseIgG2b IF,IHC,IP,WB Purified 150µg 610108
PKCd Hu,Ms,Rat 14 MouseIgG2b IF,IHC,WB Purified 150µg 610398
PKCe Chick,Dog,Hu,Ms,Rat 21 MouseIgG2a IF,IHC,IP,WB Purified 150µg 610086
PKCl Chick,Dog,Hu,Ms,Rat 41 MouseIgG1 IF,IHC,IP,WB Purified 150µg 610208
PKCq Hu 27 MouseIgG2a IF,IP,WB Purified 150µg 610090
PKCb Chick,Hu,Ms,Rat 36 MouseIgG2b IF,IHC,IP,WB Purified 50µg 610127
IF,IHC,IP,WB Purified 150µg 610128
bdbiosciences.com35Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Kinases and Phosphatases (continued)
description react clone isotype apps format size cat.no.
PKCg Ms,Rat 20 MouseIgG1 IF,WB Purified 50µg 611158
PKCiota Chick,Dog,Hu,Ms,Rat 23 MouseIgG2b IF,IHC,IP,WB Purified 50µg 610175
IF,IHC,IP,WB Purified 150µg 610176
Dros 23 MouseIgG2b IF,IHC,IP,WB Purified 50µg 612645
PP2A Catalytic a Dog,Hu,Ms,Rat 46 MouseIgG1 IF,IHC,IP,WB Purified 150µg 610556
PRK1 (PKN) Dog,Hu,Ms,Rat 49 MouseIgG1 IF,IP,WB Purified 150µg 610687
PYK2 (CAK b) Chick,Dog,Frog,Hu,Ms,Rat 11 MouseIgG1 IF,IHC,IP,WB Purified 150µg 610549
Synaptojanin 1 Ms,Rat 26 MouseIgG1 WB Purified 50µg 612248
WB Purified 150µg 612249
TAO1 Hu,Ms,Rat 22 MouseIgG2a IF,WB Purified 150µg 611369
Yes Chick,Dog,Hu,Ms,Rat 1 MouseIgG1 IF,IHC,WB Purified 150µg 610376
bdbiosciences.com36 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Functional Assays and Discovery Reagents (continued)
Phospho-specific Antibodies
description react clone isotype apps format size cat.no.
aSynuclein (PY125) Hu I57-628 MsIgG2b,k WB Purified 100µg 558246
b-Dystroglycan (pY892) Hu 27.1 MsIgG1 IF,IHC,WB Purified 50µg 612524
IF,IHC,WB Purified 150µg 612525
Actopaxin (pS8) Hu J160-366 MsIgG1,k WB Purified 100µg 558374
Akt (pS472/pS473) (PKBa) Hu,Ms,Rat 104A282 MsIgG1 IP,WB Purified 50µg 550747
Akt (pS473) Hu J177-204.20 MsIgG1,k WB Purified 100µg 558368
Akt (pT308) Hu,Ms J1-223.371 MsIgG1,k WB Purified 100µg 558316
c-Cbl (pY700) Hu 47/c-Cbl(Y700) MsIgG1 IF,IHC,WB Purified 50µg 612304
IF,IHC,WB Purified 150µg 612305
c-Cbl (pY774) Hu 29/c-Cbl(Y774) MsIgG1 WB Purified 100µg 558035
c-Jun (pS63) Hu 2 MsIgG1 IHC(F),WB Purified 100µg 558036
Caveolin (pY14) Hu,Ms,Rat 56 MsIgG1 IF,IHC,WB Purified 50µg 611338
IF,IHC,WB Purified 150µg 611339
Caveolin 2 (pY27) Hu 40/Caveolin2 MsIgG1,k WB Purified 100µg 558364
CD22 (BL-CAM) (pY828) Hu 46 MsIgG1 IHC,WB Purified 100µg 558029
CD22 (BL-CAM) (pY843) (BL-CAM) Hu 12a MsIgG1 IHC,WB Purified 100µg 558030
CD117 (pY568/pY570) (c-Kit) Hu K39-686 MsIgG1,k WB Purified 100µg 558390
CD221 (pY950) (IGF-1 Receptor) Hu J95-626 MsIgG2b,k WB Purified 100µg 558373
Cdk1/Cdc2 (pY15) Hu 44 MsIgG1 IHC,WB Purified 50µg 612306
IHC,WB Purified 150µg 612307
CREB (pS133) Hu J151-21 MsIgG1,k WB Purified 100µg 558359
CrkL (pY207) Hu K30-391.11.30 MsIgG2a,k WB Purified 100µg 558386
EGF Receptor (pY845) Hu 12A3 MsIgG1,k WB Purified 100µg 558381
EGF Receptor (pY1173) Hu 9H2 MsIgG1,k WB Purified 100µg 558382
EGF Receptor, Activated Form Hu 74 MsIgG1 IF,IP,WB Purified 50µg 610025
IF,IP,WB Purified 150µg 610026
ERK1/2 (pT202/pY204) (p44/42 MAPK) Hu,Ms,Rat 20A MsIgG1 IHC,WB Purified 50µg 612358
IHC,WB Purified 150µg 612359
A key area of neuroscience is the study of intracellular communi-
cation. Binding of ligands to receptors initiates a phosphorylation
signal cascade mediated via kinases and phosphatases (localized
in the cell membrane, cytoplasm, and nucleus). Phosphorylation of
tyrosine, serine, and threonine residues is critical for the control of
protein activity involved in cellular events. A myriad of kinases and
phosphatases regulates intracellular protein phosphorylation in many
different cell signaling pathways, such as those regulating apoptosis,
growth and cell cycle control, neurotransmitter regulation, and stress
responses.
Historically, phosphorylation detection has been performed using
techniques such as radiometric kinase assays and phosphoamine
acid labeling. In recent years, mass spectrometry techniques have
also been developed, but have a tendency to be time consuming
and cumbersome. However, the advent of phospho-specific anti-
bodies has facilitated the use of more straightforward techniques
such as WB, IP, and IF. BD Biosciences, in collaboration with Garry
Nolan, PhD at Standford University, has developed the
BD™ Phosflow technique. This technology combines phospho-
specific antibodies with the power of flow cytometry to enhance
phosphoprotein studies by reducing sample size and increasing
quantitative analysis, while allowing multiparameter analysis of
single cells and distinct cell subpopulations. This approach has been
extended using BD™ Cytometric Bead Arrays (CBA) to analyze
protein phosphorylation in cell and tissue lysates. BD Biosciences
offers a variety of phospho-specific antibodies for a variety of experi-
mental techniques, including western blotting, immunoparticipation,
immunohistochemistry, immunofluorescence microscopy, and flow
cytometry.
The reagents, instruments, and tools that BD Biosciences offers are
designed to facilitate investigation of the signal transduction and
apoptotic pathways linking cellular components and processes to
one another during development, homeostasis, and disease.
Spotlight on Phoshoprotein Analysis
bdbiosciences.com37Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Phospho-specific Antibodies (continued)
description react clone isotype apps format size cat.no.
Ezrin (pT567) Hu J37-954.281.307 MsIgG1,k WB Purified 100µg 558357
Ezrin (pY353) Hu I66-386 MsIgG1 IHC,WB Purified 100µg 558033
FADD (pS194) Hu J119-857.36 MsIgG1,k WB Purified 100µg 558370
FAK (pY397) Hu,Ms 18 MsIgG1 IF,WB Purified 50µg 611806
IF,WB Purified 150µg 611807
FAK (pY397) Hu 14 MsIgG1 IF,IHC,WB Purified 50µg 611722
IF,IHC,WB Purified 150µg 611723
Fyn (pY528)/c-Src (pY530) Hu 31 MsIgG2b IHC,WB Purified 50µg 612668
IHC,WB Purified 150µg 612669
GSK-3b (pY216) Hu,Ms,Rat 13a MsIgG1 IHC,WB Purified 50µg 612312
IHC,WB Purified 150µg 612313
IkBa (pS32/pS36) Hu 39A1431 MsIgG1 WB Purified 50µg 551818
IRS-1 (pY896) Hu K9-211 MsIgG2a,k WB Purified 100µg 558378
JNK (pT183/pY185) Hu 41 MsIgG1 IHC,WB Purified 50µg 612540
IHC,WB Purified 150µg 612541
MARCKS (pS152/pS156) Ms I84-1233 MsIgG1,k WB Purified 100µg 558380
MEK1 (pS298) Hu,Ms J114-64 MsIgG1,k WB Purified 100µg 558375
Neurofilament NF-H Rat RNF405 MsIgM WB Purified 50µg 551958
RNF404 MsIgG2a WB Purified 50µg 551348
Neurofilament NF-M Rat RNF406 MsIgG1 WB Purified 50µg 551962
NF-kB (pS529) Hu K10-895.12.50 MsIgG2b,k WB Purified 100µg 558393
NF-kB p65 (pS536) Hu J144-460 MsIgG1,k WB Purified 100µg 558377
p38 MAPK (pT180/pY182) Hu,Ms,Rat 36 MsIgG1 IF,IHC,WB Purified 50µg 612288
IF,IHC,WB Purified 150µg 612289
30 MsIgG1 IF,IHC,WB Purified 50µg 612280
IF,IHC,WB Purified 150µg 612281
p53 (pS37) Hu J159-641.15.4 MsIgG1,k WB Purified 100µg 558369
p53 (pS46) Hu I117-1091 MsIgG1,k IHC,WB Purified 100µg 558245
p90 RSK1 (pS380) Hu 20a MsIgG1 IHC,WB Purified 50µg 612692
IHC,WB Purified 150µg 612693
p120 Catenin (pY228) Hu 21a MsIgG1 IF,IHC,WB Purified 50µg 612536
IF,IHC,WB Purified 150µg 612537
p130Cas (pY249) Hu J169-757.12.2 MsIgG2b,k WB Purified 100µg 558401
Paxillin (pY31) Hu 19/Paxillin(Y31) MsIgG1,k WB Purified 100µg 558356
Paxillin (pY118) Hu,Ms 30 MsIgG1 IHC,WB Purified 50µg 611724
IHC,WB Purified 150µg 611725
PDPK1 (pS241) (PDK1) Hu J66-653.44.22 MsIgG1,k WB Purified 100µg 558395
PLK1 (pT210) Hu K50-483 MsIgG1,k WB Purified 100µg 558400
Progesterone Receptor (pS190) Hu 1154/F12 MsIgG1,k WB Purified 100µg 558387
Rb (pS780) Hu J146-35 MsIgG1,k IHC,WB Purified 100µg 558385
Rb (pS807/pS811) Hu J112-906 MsIgG1,k WB Purified 100µg 558389
Rb, Underphosphorylated (a.a. 514-610) Hu G99-549 MsIgG1 IHC,IP,WB Purified 100µg 554164
Stat1 (pY701) Hu,Ms 14 MsIgG1 IF,IP,WB Purified 50µg 612132
IF,IP,WB Purified 150µg 612133
4a MsIgG2a IF,IHC,IP,WB Purified 50µg 612232
IF,IHC,IP,WB Purified 150µg 612233
Stat2 (pY690) Hu 7a/Stat2(pY690) MsIgG1,k IHC,WB Purified 100µg 558095
bdbiosciences.com38 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Functional Assays and Discovery Reagents (continued)
Phospho-specific Antibodies (continued)
description react clone isotype apps format size cat.no.
Stat3 (pS727) Hu 49 MsIgG1 IF,IHC,WB Purified 50µg 612542
IF,IHC,WB Purified 150µg 612543
Stat3 (pY705) Hu,Ms 4 MsIgG2a IHC,WB Purified 50µg 612356
IHC,WB Purified 150µg 612357
Stat4 (pY693) Hu 38/p-Stat4 MsIgG2b IHC,WB Purified 50µg 612738
IHC,WB Purified 150µg 612739
Stat5 (pY694) Hu Polyclonal RabIg IF,WB Purified 50µg 611818
IF,WB Purified 150µg 611819
47 MsIgG1 IHC,WB Purified 50µg 611964
IHC,WB Purified 150µg 611965
Stat6 (pY641) Hu Polyclonal RabIg IF,WB Purified 50µg 611820
IF,WB Purified 150µg 611821
18 MsIgG2a IF,IHC,WB Purified 50µg 611566
IF,IHC,WB Purified 150µg 611567
Ms J71-773.58.11 MsIgG1,k WB Purified 100µg 558241
Tyk2 (pY1054/pY1055) Hu I114-617 MsIgG1,k WB Purified 100µg 558394
bdbiosciences.com39Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
BD Phosflow Reagents*
description clone apps format size cat.no.
AKT (pS473) F29-763 IC/FCM AlexaFluor®647 50tests 558314
AKT (pT308) J1-223.371 IC/FCM PE 50tests 558275
BLNK (pY84) J117-1278 IC/FCM AlexaFluor®488 50tests 558444
IC/FCM AlexaFluor®647 50tests 558443
IC/FCM PE 50tests 558442
Btk (pY551) & Itk (pY511) 24a/BTK(Y551) IC/FCM PE 50tests 558129
IC/FCM AlexaFluor®488 50tests 558130
IC/FCM AlexaFluor®647 50tests 558134
c-Cbl (pY700) 47/c-Cbl(Y700) IC/FCM PE 50tests 558087
IC/FCM AlexaFluor®488 50tests 558101
IC/FCM AlexaFluor®647 50tests 558100
c-Cbl (pY774) 29/c-Cbl(Y774) IC/FCM PE 50tests 558102
IC/FCM AlexaFluor®647 50tests 558103
Caveolin (pY14) 56 IC/FCM PE 50tests 612568
CD140b (pY771) J23-1044 IC/FCM PE 50tests 558426
IC/FCM AlexaFluor®488 50tests 558424
IC/FCM AlexaFluor®647 50tests 558425
CD140b (pY857) J24-618 IC/FCM PE 50tests 558429
IC/FCM AlexaFluor®488 50tests 558427
IC/FCM AlexaFluor®647 50tests 558428
CD140b (pY1009) J25-602 IC/FCM PE 50tests 558322
IC/FCM AlexaFluor®647 50tests 558323
CD140b (pY1021) J105-412 IC/FCM PE 50tests 558417
IC/FCM AlexaFluor®488 50tests 558419
IC/FCM AlexaFluor®647 50tests 558418
CD247 (pY142) K25-407.69 IC/FCM PE 50tests 558448
IC/FCM AlexaFluor®488 50tests 558486
IC/FCM AlexaFluor®647 50tests 558489
CREB (pS133) J151-21 IC/FCM PE 50tests 558436
IC/FCM AlexaFluor®488 50tests 558435
IC/FCM AlexaFluor®647 50tests 558434
EGF Receptor (pY845) 12A3 IC/FCM AlexaFluor®647 50tests 558523
EGF Receptor (pY1173) 9H2 IC/FCM AlexaFluor®647 50tests 558524
BD Phosflow is a highly innovative flow cytometry based technology,
enabling activation state analysis of multiple proteins at single-cell levels.
Using phosphorylation site specific antibodies in combination with
cell surface markers, you can study phosphorylation events directly in
small subpopulations of complex primary samples.
This revolutionary technology will not only enhance cell signaling
network analysis and high content secondary screening for drug
discovery and development, but also will enable the identification of
signaling pathway dependent biosignatures for disease.
BD Phosflow Reagents BD Phosflow Advantages
• Collect data for individual cells
• Analyze rare cell types directly in complex primary samples
• Correlate multiple phosphorylation events simultaneously
• Perform rapid and scalable screens in 96-well plates using
BD FACS™ flow cytometers with the BD™ HTS option
bdbiosciences.com40 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
*Visit bdbiosciences .com/phosflow for a complete listing of BD Phosflow reagents.
Functional Assays and Discovery Reagents (continued)
BD Phosflow Reagents (continued)
description clone apps format size cat.no.
ERK1/2 (pT202/pY204) 20A IC/FCM PE 50tests 612566
IC/FCM AlexaFluor®488 50tests 612592
IC/FCM AlexaFluor®647 50tests 612593
ERK2 G263-7 IC/FCM AlexaFluor®488 50tests 558525
IC/FCM AlexaFluor®647 50tests 558526
FAK (pS910) K73-480 IC/FCM PE 50tests 558540
IC/FCM AlexaFluor®488 50tests 558544
Histone H3 (pS28) HTA28 IC/FCM AlexaFluor®647 50tests 558217
IRS-1 (pY896) K9-211 IC/FCM PE 50tests 558440
IC/FCM AlexaFluor®647 50tests 558441
JNK (pT183/pY185) Polyclonal IC/FCM,IHC(F),WB Purified 50tests 558268
LAT (pY171) I58-1169 IC/FCM PE 50tests 558487
IC/FCM AlexaFluor®488 50tests 558519
IC/FCM AlexaFluor®647 50tests 558518
LAT (pY226) J96-1238.58.93 IC/FCM PE 50tests 558433
IC/FCM AlexaFluor®488 50tests 558430
IC/FCM AlexaFluor®647 50tests 558432
Lck (Y505) 4/Lck-Y505 IC/FCM PE 50tests 558552
IC/FCM AlexaFluor®488 50tests 557879
MEK1/2 (pS222) Polyclonal IC/FCM,IHC(F), Purified 50tests 558281 IHC(F)†,WB
NF-kB p65 (pS529) K10-895.12.50 IC/FCM PE 50tests 558423
IC/FCM AlexaFluor®488 50tests 558421
IC/FCM AlexaFluor®647 50tests 558422
Notch1 mN1A FCM,IC/FCM,WB PE 100µg 552768
NPM-ALK ALK1 IC/FCM FITCSet 100tests 559818
IC/FCM PESet 100tests 559257
p38 MAPK (pT180/pY182) 36 IC/FCM PE 50tests 612565
IC/FCM AlexaFluor®488 50tests 612594
IC/FCM AlexaFluor®647 50tests 612595
p120 Catenin (pS288) 17/catenin IC/FCM AlexaFluor®647 50tests 558561
p130Cas (pY249) J169-757.12.2 IC/FCM AlexaFluor®488 50tests 558546
IC/FCM AlexaFluor®647 50tests 558547
PLC-g1 (Y783) 27 IC/FCM AlexaFluor®488 50tests 557884
IC/FCM AlexaFluor®647 50tests 557883
PLC-g2 (pY759) K86-689.37 IC/FCM PE 50tests 558490
IC/FCM AlexaFluor®488 50tests 558507
IC/FCM AlexaFluor®647 50tests 558498
PLK1 (pT210) K50-483 IC/FCM PE 50tests 558445
IC/FCM AlexaFluor®488 50tests 558446
IC/FCM AlexaFluor®647 50tests 558447
Rb (a.a. 332-344) G3-245 IC/FCM FITCSet 100tests 556538
IC/FCM PESet 100tests 556539
Rb (pS780) J146-35 IC/FCM AlexaFluor®488 50tests 558554
IC/FCM AlexaFluor®647 50tests 558555
Rb, underphosphorylated (a.a. 514-610) G99-549 IC/FCM FITCSet 100tests 550501
IC/FCM PESet 100tests 550502
†Antigen retrieval required
*Visit bdbiosciences .com/phosflow for a complete listing of BD Phosflow reagents.
bdbiosciences.com41Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
BD Phosflow Reagents* (continued)
description clone apps format size cat.no.
SLP-76 (pY128) J141-668.36.58 IC/FCM PE 50tests 558437
IC/FCM AlexaFluor®488 50tests 558439
IC/FCM AlexaFluor®647 50tests 558438
Stat1 (pY701) 4a IC/FCM PE 50tests 612564
IC/FCM AlexaFluor®488 50tests 612596
IC/FCM AlexaFluor®647 50tests 612597
Stat3 (pS727) 49/p-Stat3 IC/FCM AlexaFluor®488 50tests 558085
IC/FCM AlexaFluor®647 50tests 558099
Stat3 (pY705) 4 IC/FCM PE 50tests 612569
FCM,WB AlexaFluor®488 50tests 557814
FCM,WB AlexaFluor®647 50tests 557815
Stat4 (pY693) 38/p-Stat4 IC/FCM PE 50tests 558249
IC/FCM AlexaFluor®488 50tests 558136
IC/FCM AlexaFluor®647 50tests 558137
Stat5 (pY694) 47 IC/FCM PE 50tests 612567
IC/FCM AlexaFluor®488 50tests 612598
IC/FCM AlexaFluor®647 50tests 612599
Stat6 (pY641) 18 IC/FCM PE 50tests 612701
IC/FCM AlexaFluor®488 50tests 612600
IC/FCM AlexaFluor®647 50tests 612601
J71-773.58.11 IC/FCM PE 50tests 558252
IC/FCM AlexaFluor®488 50tests 558243
IC/FCM AlexaFluor®647 50tests 558242
Syk (p352) Pan/Phospho 4D10 IC/FCM FITC 50tests 557952
Syk (pY348) I120-722 IC/FCM PE 20µL 558529
TCL1 H29-1349 IC/FCM PE 50tests 558025
Zap70 (pY292) J34-602 IC/FCM PE 50tests 558510
IC/FCM AlexaFluor®488 50tests 558516
IC/FCM AlexaFluor®647 50tests 558515
Zap70 (Y319)/Syk (Y352) 17a IC/FCM PE 50tests 557881
IC/FCM AlexaFluor®488 50tests 557818
IC/FCM AlexaFluor®647 50tests 557817
BD Phosflow Supporting Reagents
description clone isotype format size cat.no.
BD Cytofix Fixation Buffer 100mL 554655
BD Phosflow Fix Buffer 250mL 557870
BD Phosflow Lyse/Fix Buffer (5X) 250mL 558049
BD Phosflow Perm/Wash Buffer I 125mL 557885
BD Phosflow Perm Buffer II 125mL 558052
BD Phosflow Perm Buffer III 125mL 558050
Staining Buffer (FBS) 500mL 554656
Mouse IgG1 Isotype Control MOPC-21 MsIgG1 AlexaFluor®488 50tests 557782
AlexaFluor®647 50tests 557783
PE 50tests 551436
Mouse IgG2a Isotype Control MOPC-173 MsIgG2a PerCP-Cy™5.5 50tests 558020
*Visit bdbiosciences .com/phosflow for a complete listing of BD Phosflow reagents.
bdbiosciences.com42 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Functional Assays and Discovery Reagents (continued)
Cytoskeletal Proteins
description react clone isotype apps format size cat.no.
4.1N Chick,Dog,Hu,Ms,Rat 4 MouseIgG1 IF,WB Purified 50µg 611836
Actin Chick 4 MouseIgG1 IF,WB Purified 50µg 612656
IF,WB Purified 150µg 612657
Arp3 Chick,Dog,Hu,Ms,Rat 4 MouseIgG1 IF,WB Purified 50µg 612134
150µg 612135
BART Hu,Ms,Rat 6 MouseIgG1 WB Purified 50µg 612036
WB Purified 150µg 612037
CLIP-115 Hu,Ms,Rat 14 MouseIgG1 IF,WB Purified 50µg 611142
IF,WB Purified 150µg 611143
Cofilin Dog,Hu,Ms,Rat 32 MouseIgG1 WB Purified 50µg 612144
150µg 612145
Collybistin Ms,Rat 3 MouseIgG1 IF,WB Purified 50µg 612076
Contactin Hu,Ms,Rab,Rat 41 MouseIgG1 IF,IP,WB Purified 50µg 610578
IF,IP,WB Purified 150µg 610579
Dystrobrevin Ms,Rab,Rat 23 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610766
Rab,Rat 38 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610780
IF,IHC,IP,WB Purified 150µg 610781
The cytoskeleton regulates cell shape,
transport, motility, and integrity.
Cytoskeleton consists of filament systems
with a large number of associated proteins
that regulate each system. Cytoskeletal
filaments that are found in all cells can be
grouped into microfilaments, intermediate
filaments, and microtubules.
Microfilaments Microfilaments are the smallest (7–9 nm in diameter) of the three cytoskeletal filaments. They consist of actin and tropomyosin in association with a large number of other proteins.
• Mammalian actin exists in six different forms (isoforms): four associated with different types of muscle and two cytoskeletal isoforms found in all non-muscle cells.
- cytoskeletal isoforms are b and g- muscle isoforms are a (skeletal), b
(cardiac), g (smooth), and g (enteric)
• Actin is usually the most abundant protein in cells.
• It is 375 amino acids in length and isoforms can be separated on two-dimensional protein gels.
• b and g cytoskeletal actin differ by four amino acids at their amino end.
• Actin can be identified in its monomer (g-actin, globular) or polymer (f-actin, filamentous) form.
• Actin can also be identified by antibodies to all, or specific, isoforms.
• Actin mRNA is localized to specific regions within the cell cytoplasm.
Intermediate FilamentsIntermediate filaments are rope-like fibers made of eight tetramers, 10 nm in diameter. They are intermediate in size between microfilaments and microtubules. Different cell types express different combinations of intermediate filaments.
• Their main function is to maintain cell integrity. Most cells express more than one intermediate filament protein.
• The nucleus has its ownintermediate filament network (lamins).
• Some examples are keratin (skin),neurofilament (neurons), and glial fibrillary acid protein (GFAP, glia).
MicrotubulesLargest of the three filaments, 24 nm in diameter, microtubules form hollow tubes made of 13 parallel proto-filaments, involved with cytoplasmic transport of organelles, proteins, and vesicles.
• Different motors move in different directions on microtubules.
• Tubules composed of heterodimers of tubulin. There are two isoforms of tubulin: b and g.
• Stabilized by a large family of different microtubule associated proteins (MAPs). The most famous MAP is tau, found in neurofibrillary tangles associated with Alzheimer’s Disease (AD).78
Cytoskeletal Proteins
bdbiosciences.com43Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Cytoskeletal Proteins (continued)
description react clone isotype apps format size cat.no.
GFAP Bov,Chick,Dog,GPig, 1B4 MouseIgG2b IF,IHC(F),WB Purified 500µg 556328 Hu,Ms,Pig,Rab,Rat,Sheep
2E1 MouseIgG2b IF,IHC(F),WB Purified 500µg 556329
4A11 MouseIgG2b IF,IHC(F),WB Purified 500µg 556327
Chick,Hu,Ms,Rat 52 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610565
IF,IHC,IP,WB Purified 150µg 610566
GFAP Cocktail Bov,Chick,Dog,GPig, 4A11,1B4,2E1 MouseIgG2b IF,IHC(F),WB Purified 500µg 556330 Hu,Ms,Pig,Rab,Rat,Sheep
KAP3A Chick,Dog,Hu,Ms,Rat 14 MouseIgG1 IF,IP,WB Purified 50µg 610637
Katanin p80 Hu 19 MouseIgG1 IF,WB Purified 150µg 611411
KIF1A Ms 16 MouseIgG1 IF,WB Purified 50µg 612094
KIF3A Hu,Ms,Rat 28 MouseIgG1 IF,WB Purified 150µg 611509
Laminin B2 Hu,Rat 22 MouseIgG1 IF,IP,WB Purified 50µg 610722
MAP1B Ms 6 MouseIgG2a WB Purified 50µg 612678
WB Purified 150µg 612679
MAP1B (LC1) Ms 19 MouseIgG1 WB Purified 50µg 612680
WB Purified 150µg 612681
MAP2 Bov,Frog,Hu,Qua,Rat Ap20 MouseIgG1 IHC(Fr),IP,WB Purified 100µg 556320
MAP2B Hu,Ms,Rat 18 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610460
MKLP1 Hu 24 MouseIgG2a WB Purified 50µg 612682
WB Purified 150µg 612683
Nestin Hu,Ms,Rat 25 MouseIgG1,k IF,WB Purified 50µg 611658
IF,WB Purified 150µg 611659
Rat Rat401 MouseIgG1 EM,FCM,IF, Purified 100µg 556309 IHC(P),WB
Neurofilament NF-H Rat RNF402 MouseIgM WB Purified 50µg 551349
RNF404 MouseIgG2a WB Purified 50µg 551348
RNF405 MouseIgM WB Purified 50µg 551958
Neurofilament NF-M Rat RNF403 MouseIgG1 WB Purified 50µg 551957
RNF406 MouseIgG1 WB Purified 50µg 551962
Ninjurin Dog,Hu,Ms,Rat 50 MouseIgG2a IF,IHC,WB Purified 50µg 610776
150µg 610777
Occludin Dog,Hu,Ms,Rat 19 MouseIgG1 WB Purified 50µg 611090
150µg 611091
p150 Glued Chick,Dog,Hu,Ms,Rat 1 MouseIgG1 IF,IP,WB Purified 50µg 610473
IF,IP,WB Purified 150µg 610474
Rat 12 MouseIgG1 IF,WB Purified 50µg 612708
IF,WB Purified 150µg 612709
p24 Ms,Rat 22 MouseIgG1 IF,WB Purified 50µg 611616
IF,WB Purified 150µg 611617
Tau Rat 15 MouseIgG2b IF,IHC,IP,WB Purified 50µg 610672
IF,IHC,IP,WB Purified 150µg 610673
TAU-5 MouseIgG1 IHC(F),WB Purified 100µg 556319
b-Tubulin Bov,Hu,Ms,Rat 5H1 MouseIgM ICC,WB Purified 100µg 556321
VAP33 Dog,Hu,Ms,Rat 8 MouseIgM IF,WB Purified 50µg 612180
bdbiosciences.com44 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Functional Assays and Discovery Reagents (continued)
bdbiosciences.com45Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Development and Differentiation
description react clone isotype apps format size cat.no.
d-Catenin Ms,Rat 30 MouseIgG1 WB Purified 50µg 611536
150µg 611537
ADNP Hu 45 MouseIgG1 IF,WB Purified 50µg 612512
BERP Ms,Rat 27 MouseIgG1 IF,WB Purified 50µg 611732
IF,WB Purified 150µg 611733
beta -Dystroglycan Hu 56 MouseIgG2a IF,WB Purified 50µg 612090
IF,WB Purified 150µg 612091
beta -Dystroglycan (pY892) Hu 27.1 MouseIgG1 FCM,IF,WB Purified 50µg 612524
FCM,IF,WB Purified 150µg 612525
Brevican Dog,Ms,Rat 2 MouseIgG1 IF,WB Purified 50µg 610894
IF,WB Purified 150µg 610895
CD171 Hu 5G3 MouseIgG2a IP Purified 100µg 554273
CD56 Ms 12F8 RatIgM IHC(Fr),WB Purified 100µg 556325
12F11 RatIgG2a IHC(F),IHC(Fr), Purified 100µg 556323 WB,IHC(P)
Ms,Rat N-CAM13 MouseIgG2a IHC(Fr),WB Purified 100µg 556324
CRMP5 Ms 8 MouseIgG1 IF,WB Purified 50µg 612364
Doublecortin Ms 30 MouseIgG1 IF,WB Purified 50µg 611706
DP103/Gemin3 Hu 2 MouseIgG2b WB Purified 50µg 612152
150µg 612153
ENC-1 Dros 7 MouseIgG1 IF,IHC,WB Purified 50µg 612628
Hu,Ms,Rat 7 MouseIgG1 IF,WB Purified 50µg 611210
FAK Chick,Dog,Hu,Ms,Rat 77/FAK MouseIgG1 IF,IHC,IP,WB Purified 50µg 610087
150µg 610088
FAK (pY397) Hu,Ms 14 MouseIgG1 IF,WB Purified 150µg 611723
18 MouseIgG1 IF,WB Purified 150µg 611807
Fyn Dog,Hu,Rat 25 MouseIgG2b IF,IHC,IP,WB Purified 50µg 610163
IF,IHC,IP,WB Purified 150µg 610164
Fyn (pY528)/c-Src (pY530) Hu 31 MouseIgG2b WB Purified 50µg 612668
WB Purified 150µg 612669
Galpha o Hu 23 MouseIgG2a IF,WB Purified 50µg 612526
IF,WB Purified 150µg 612527
For most species, the nervous system is the most complex part of
the organism. For instance, in mammalian species, over one third of
the genome is expressed in the nervous system. The development
process is complex and requires the precise control of many events
that generate a large variety of cell types with absolute correct
positioning and interconnections. The development of the nervous
system starts early in embryonic life and creates and maintains the
complex system of structures—which includes the brain, spinal
cord, and peripheral nerves—throughout the life of the individual.
Very early in the embryonic life, the neural plate and neural tube
are induced and the neural precursors form and proliferate. These
precursors will develop into neurons or glial cells. Neuronal and
glial precursors migrate from their site of origin to final positions in
the region they will occupy. They will then differentiate into mature
phenotypes, expressing appropriate functional molecules, extending
axons, setting up synaptic connections, and undergoing myelination
and additional modifications to delineate mature structures. During
the process, many neurons undergo programmed cell death.
Some key processes and molecules that guide development are:
• The effects of morphogens and Hox genes in pattern formation and the specification of cell fate
• Cell-to-cell interactions that regulate specification, and the molecules that effect those, such as notch and delta interactions
• Boundary regions that contain specialized cells such as “organizers”
• Growth in cortex regulated and guided by glial cells
• The effects of neurotransmitters
• The role of genetic programs in the formation of synapses versus activity-regulated remodeling of neuronal connections
Development and Differentiation
Development and Differentiation (continued)
description react clone isotype apps format size cat.no.
GRIFIN Rat 50 MouseIgG1 IF,WB Purified 50µg 611502
IF,WB Purified 150µg 611503
HERC2 Ms 17 MouseIgG1 WB Purified 50µg 612366
Jagged1 Hu 21 MouseIgG1 WB Purified 50µg 612346
Janusin Hu,Ms,Rat 9 MouseIgG1 IF,IP,WB Purified 50µg 610629
KNP-1 (HES1) Chick,Dog,Hu,Ms,Rat 35 MouseIgG1 IF,WB Purified 50µg 611596
150µg 611597
Lamin A/C Chick,Dog,Hu,Ms,Rat 14 MouseIgG1 IF,WB Purified 50µg 612162
IF,WB Purified 150µg 612163
LAR Bov,Hu,Ms,Rat 7 MouseIgG2a IF,IP,WB Purified 50µg 610350
IF,IP,WB Purified 150µg 610351
LR11 Hu,Ms,Rat 48 MouseIgG2a IF,WB Purified 50µg 611860
IF,WB Purified 150µg 611861
Dros 48 MouseIgG2a IF,IHC,WB Purified 50µg 612633
MASH1 Ms,Rat 24B7.2D11 MouseIgG1 WB Purified 100µg 556604
M-Cadherin Ms,Rat 5 MouseIgG2a IF,WB Purified 50µg 611100
150µg 611101
Mena Chick,Hu,Ms,Rat 21 MouseIgA IF,WB Purified 50µg 610692
150µg 610693
Myelin Basic Protein Gt,Ms,Prim,Rab,Rat,Sheep MyelinBP MouseIgG2b WB Purified 100µg 559904
N-Cadherin Chick,Hu,Ms,Rat 32 MouseIgG1 IF,WB Purified 50µg 610920
150µg 610921
Neurogenin 3 Hu,Ms,Rat 7 MouseIgG1 IF,IP,WB Purified 50µg 610790
Neuroglycan C Rat 1 MouseIgG1 IF,WB Purified 50µg 610986
Neuropilin-2 Rat 54 MouseIgG1 IF,WB Purified 50µg 611262
Nogo-A Rat 17 MouseIgG1 IF,WB Purified 50µg 612238
Pax-5 Hu,Ms 24 MouseIgG1 IF,WB Purified 50µg 610862
150µg 610863
Pit-1 Rat 29 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610223
R-Cadherin Ms,Rat 48 MouseIgG1 IP,WB Purified 50µg 610414
150µg 610415
Sema4C Hu 37 MouseIgG1 WB Purified 50µg 612486
TOAD-64 Ms,Rat Polyclonal RabbitIg WB Purified 100µL 551357
Tubby Ms 40 MouseIgG1 WB Purified 50µg 612106
V-1 Hu,Ms,Rat 49 MouseIgG2b IF,WB Purified 50µg 611830
IF,WB Purified 150µg 611831
ZPR1 Ms 8 MouseIgG1 WB Purified 150µg 612129
bdbiosciences.com46 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Functional Assays and Discovery Reagents (continued)
Growth Factor Related
description react clone isotype apps format size cat.no.
B2 Bradykinin Receptor Chick,Dog,Hu,Ms,Rat 20 MouseIgG2b IF,IHC,IP,WB Purified 50µg 610451
MouseIgG2b IF,IHC,IP,WB Purified 150µg 610452
c-erbB-2 Hu 42 MouseIgG2b IF,IHC,IP,WB Purified 50µg 610161
150µg 612162
Endopeptidase 3.4.24.16 Hu,Ms,Rat 35 MouseIgA WB Purified 50µg 611240
WB Purified 150µg 611241
Glucocorticoid Receptor Hu 41 MouseIgG1 IF,WB Purified 50µg 611226
IF,WB Purified 150µg 611227
IL-6 Hu MQ2-13A5 RatIgG1 FCM,IC Purified 100µg 554542
Neu NA/LE 500µg 554541
LEDGF Hu 26 MouseIgG1 IF,WB Purified 50µg 611714
IF,WB Purified 150µg 611715
Neurotensin Receptor 3 Hu 48 MouseIgG1 IF,WB Purified 50µg 612100
IF,WB Purified 150µg 612101
Thrombin Receptor (PAR1) Hu,Ms 14 MouseIgG1 IF,WB Purified 50µg 611522
150µg 611523
TrkB Ms,Rat 47 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610101
IF,IHC,IP,WB Purified 150µg 610102
Polyclonal RabbitIg IF,WB Purified 50µg 611640
IF,WB Purified 150µg 611641
VEGF Hu G153-694 MouseIgG2b IF,IHC(F) Purified 500µg 555036
G143-850 MouseIgG2a IP,WB Purified 500µg 554539
A principal class of intercellular signaling molecules is the protein
growth factors. Neurotrophins and the neurotrophin receptors
represent a family of molecules that regulate growth and survival in
the nervous system. The first functional neurotrophin described is
nerve growth factor (NGF). NGF was discovered 50 years ago as a
molecule that promoted the survival and differentiation of sensory
and sympathetic neurons. NGF also affects additional processes such
as differentiation and microtubule assembly in some cells. Other
neurotrophin family members include brain-derived neurotrophic
factor (BDNF), neurotrophin-3 (NT3), and NT4/5. Neurotrophins have
specific patterns of expression and affect different and overlapping
neuronal populations. Other molecules that have been shown to
play roles in proliferation are GDNF (glial cell line derived neuro-
trophic factor), CNTF (ciliary neurotrophic factor), TGF-b, LIF, IL-6,
VEGF, and FGF. Different growth factors stimulate different and over-
lapping cell types in the nervous system. TGF-b is related to injury
response and produced by both glia and neurons. VEGF appears to
have neuro-protective properties.
Soluble and membrane embedded neurotrophic factors bind to
specific receptors on responsive neurons. Neurotrophins bind the
Trk family of tyrosine kinase receptors (TrkA, TrkB, TrkC) and the
p75 neurotrophin receptor (p75NTR, CD271), the low affinity NFG
receptor subunit. The Trk receptors are responsible for most of the
survival and growth properties of the neurotrophins. The actions of
p75NTR fall into two categories. First, p75NTR is a Trk co-receptor
that can enhance or suppress neurotrophin-mediated activity.
Second, p75NTR can act autonomously in apoptosis or cell survival.
TrkB binds the neurotrophins NT3, NT4/5, and BDNF. Additionally,
CNTF and LIF act as growth factors and signal through gp130 and
the JAK/STAT pathway.
Growth Factor Related
bdbiosciences.com47Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Ion Channels and Transporters
description react clone isotype apps format size cat.no.
Aquaporin 2 Ms,Rat Polyclonal RabbitIg WB Purified 200µL 550648
Ca 2+ Channel Ca v1.2 Ms,Rat Polyclonal RabbitIg WB Purified 50µL 550716
IHC,WB Purified 200µL 550715
Connexin-43 Chick,Dog,Hu,Ms,Rat 2 MouseIgG1 IF,WB Purified 50µg 610061
150µg 610062
Erg2 Ms,Rat 46 MouseIgG1 IF,WB Purified 50µg 611076
GTRAP48 Rat 20 MouseIgG1 WB Purified 50µg 612672
WB Purified 150µg 612673
K+ Channel a Hu,Ms,Rat 32 MouseIgG1 IF,WB Purified 50µg 611248
IF,WB Purified 150µg 611249
K+ Channel SK3 Hu,Rat Polyclonal RabbitIg WB Purified 200µL 550654
Na+, K+ ATPase b 2 Hu,Ms,Rat 35 MouseIgG2b IF,WB Purified 50µg 610914
IF,WB Purified 150µg 610915
Na+, K+ ATPase b 3 Hu 46 MouseIgG1 IF,WB Purified 50µg 610992
IF,WB Purified 150µg 610993
NHE-1 Dog,Hu,Ms,Rat 54 MouseIgG1 IF,WB Purified 50µg 611774
150µg 611775
PCMT-II Bov,Dog,Hu 4 MouseIgG1 IF,IP,WB Purified 50µg 610772
PMCA2 Hu,Ms,Rat 28 MouseIgG1 IF,WB Purified 50µg 610932
Transporting molecules across a membrane can occur as either
passive transport or active transport. Passive transport does not
utilize extra energy to move molecules across the cell membrane.
Coupled transport pairs the movement of one molecule down its
gradient to move another molecule against a gradient. Examples of
passive transport are: • Gated (ligand-gated Na+ channel)
• Non-gated ion channels (K+ leak channel)
• Carriers transporters:
– Uniporter (GLUT1 uniporter)
– Symporter (Na+/glucose symporter)
– Antiporter (Na+/Ca2+ antiporter)
Transporters utilize either an electrochemical or a concentration
gradient as a source of energy. Molecules get transported down the
gradient or against the gradient for coupled transport pairs.
Active transport requires ATP to drive unfavorable movements of
molecules against their electrochemical gradient.
Examples of active transporter are Na+/K+ ATPase and ABC
transporters (such as MDR-1). They are ATP-powered pumps that
use ATP hydrolysis as an energy source, transporting against the
potential gradient. Membrane transport proteins play an important
role in establishing the membrane potential and producing action
potentials.
The membrane potential is the balance of charges between the
inside and outside of the cell and is not equal to zero. The mamma-
lian cell typically has a membrane potential of –70 millivolts inside
the cell. This negative membrane potential is due to three contribu-
tors of negative charges inside the cell: 1) K+ leak channels let
positive cations out, 2) Na+/K+ ATPase pumps three Na+ ions out of
the cell and only returns two K+ ions into the cell, and 3) protein
anions inside the cell have negative charge.
The action potential is a change in this resting potential, which
results as a way to communicate electrically within nerve cells.
The action potential is the electrical signal, but a chemical
signal exists between nerve cells. This takes place in the form of
neurotransmitters. As the action potential propagates down the
nerve axon, it depolarizes the cell membrane and opens voltage-
gated calcium channels. Calcium enters the cell at the nerve terminal
to cause vesicles to fuse with the plasma membrane and release
neurotransmitters. Once neurotransmitters enter the synaptic cleft,
calcium binds ligand-gated Na+ channels at the postsynaptic cell and
opens them. Na+ enters the postsynaptic cell to start another action
potential.
Ion Channels and Transporters
bdbiosciences.com48 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Functional Assays and Discovery Reagents (continued)
Neurotransmitter Receptor Related
description react clone isotype apps format size cat.no.
Acetylcholine Receptor a Ms,Rat 26 MouseIgG2a IF,WB Purified 50µg 610988
IF,WB Purified 150µg 610989
Acetylcholine Receptor b Hu,Rat 74 MouseIgG2b IF,IP,WB Purified 50µg 610283
IF,IP,WB Purified 150µg 610284
APM Hu,Rat 12 MouseIgG2a IF,WB Purified 50µg 611166
IF,WB Purified 150µg 611167
b Arrestin1 Hu,Ms,Rat 10 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610550
IF,IHC,IP,WB Purified 150µg 610551
Carbonic Anhydrase XIV Ms,Rat 49 MouseIgG2a WB Purified 50µg 612140
COMT Ms,Rat 4 MouseIgG1 WB Purified 50µg 611970
EAAT2 Rat 20 MouseIgG1 IF,WB Purified 50µg 611654
GABA BR2 Ms,Rat 1 MouseIgG1 IF,WB Purified 50µg 611744
GAD65 Hu,Ms,Rat,Pig GAD-6 MouseIgG2a,k IHC(Fr),IP,WB Purified 100µg 559931
GAD67 Rat 2 MouseIgG2a IF,WB Purified 50µg 611604
GluR d 2 Ms,Rab,Rat 48 MouseIgG1 IF,WB Purified 50µg 611006
GluR2 Dog,Monk,Rat 6C4 MouseIgG2a IHC(F),RIA,WB Purified 100µg 556341
GluR2/4 Hu,Monk,Rat 3A11 MouseIgG2a IHC(Fr),IP,WB Purified 100µg 556307
GluR5/6/7 (N-Terminal) Hu,Monk,Rat 4F5 MouseIgM EM,IF,IHC Ascites 250µL 556306
Glutamine Synthetase Hu,Rat 6 MouseIgG2a IF,IHC,IP,WB Purified 50µg 610517
IF,IHC,IP,WB Purified 150µg 610518
mGluR1 Ms,Rat 20 MouseIgG1 IF,WB Purified 50µg 610964
IF,WB Purified 150µg 610965
mGluR1a Rat G209-488 MouseIgG1 IHC(Fr),WB Purified 100µg 556331
G209-2048 MouseIgG1 IHC(Fr) Purified 100µg 556389
NMDAR1 Hu,Monk,Rat 54.1 MouseIgG2a EM,IHC,WB Purified 100µg 556308
NMDAR2A Ms,Rat 5 MouseIgG1 WB Purified 50µg 612286
NMDAR2B Hu,Ms,Rat 13 MouseIgG2b IF,IHC,IP,WB Purified 50µg 610416
IF,IHC,IP,WB Purified 150µg 610417
Purinergic Receptor P2X7 Rat Polyclonal RabbitIg WB Purified 50µL 550694
WB Purified 200µL 550693
Serine Racemase Ms,Rat 29 MouseIgG1 IF,WB Purified 50µg 612052
IF,WB Purified 150µg 612053
Serotonin Receptor 5-HT 2AR Hu,Monk,Rat G186-1117 MouseIgG1 IHC(Fr),WB Purified 100µg 556326
Serotonin Receptor 5-HT 2BR Hu,Rat A72-1 MouseIgG1 IHC(Fr),WB Purified 100µg 556334
Serotonin Receptor 5-HT 2CR Hu,Rat A4-2 MouseIgG1 IHC(Fr),WB Purified 100µg 556335
Substance P Pig,Rat NC1/34 RatIgG2a EM,IHC(Fr),RIA Purified 100µg 556312
Neurotransmitters are the chemical signals that relay electrical
signals between a neuron and another cell, and thus mediate many
responses within the organism. Hundreds of small molecular weight
molecules have been described as neurotransmitters. They can be
amino acids or monoamines, such as L-glutamate, GABA, dopamine,
or acetylcholine and peptides such as members of the opioid family
or hormones. Once the molecules of a neurotransmitter are released
from the synaptic vesicles of the cell, they bind to specific recep-
tors on the surface of the postsynaptic cell. Ligand-gated ionotropic
receptors are represented by receptors for acetylcholine, GABA, and
glutamate and are made up of several subunits. The metabotropic
receptors such as norepinephrine, serotonin, dopamine, and other
neuropeptides are single, transmembrane proteins that function
through GTP-binding proteins. Nerve transmissions that terminate
at a muscle cell, at the neuromuscular junction, are mediated by
acetylcholine receptors. The GABA receptor mediates the action of
g-aminobutyric acid, a major inhibitor transmitter in the brain and
spinal cord. A majority of synapses in the central nervous system
utilize glutamate as a neurotransmitter. The ionotropic receptors
glutamate receptors are subdivided into two types – receptors for
N-methyl D-aspartate (NMDAR) and non-NMDA receptors for AMPA
and kainate.
Neurotransmitter Receptor Related
bdbiosciences.com49Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Pathology
description react clone isotype apps format size cat.no.
ABR Hu,Ms,Rat 24 MouseIgG1 IF,WB Purified 50µg 611122
IF,WB Purified 150µg 611123
a-Synuclein Hu,Ms,Rat 42 MouseIgG1 IF,IP,WB Purified 50µg 610786
IF,IP,WB Purified 150µg 610787
ApoE Dog,Hu,Ms,Rat 32 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610449
IF,IHC,IP,WB Purified 150µg 610450
APP-BP1 Dog,Hu,Ms,Rat 20 MouseIgG2a IF,WB Purified 50µg 611864
IF,WB Purified 150µg 611865
Ataxin-2 Hu,Rat 22 MouseIgG1 IF,WB Purified 50µg 611378
b-Amyloid (a.a. 1-42) Hu Polyclonal RabbitIg Blot,ELISA, Purified 25µg 556501 IHC(Fr),RIA
b-Synuclein Rat 8 MouseIgG1 WB Purified 50µg 612508
Ceruloplasmin Hu,Ms,Rat 8 MouseIgG1 IF,WB Purified 50µg 611488
IF,WB Purified 150µg 611489
Choline Acetyltransferase Rat Polyclonal GoatIg IHC,WB Purified 100µL 552623
Cox-2 Chick,Hu,Ms 33 MouseIgG1 WB Biotin 50µg 610205
WB Biotin 150µg 610206
IF,IHC,IP,WB Purified 50µg 610203
IF,IHC,IP,WB Purified 150µg 610204
DDX1 Hu 22 MouseIgG1 IF,WB Purified 50µg 611206
150µg 611207
ERAB Bov,Dog,Hu,Ms,Rat 23 MouseIgG1 IF,WB Purified 50µg 611086
IF,WB Purified 150µg 611087
Fascin Ms 5 MouseIgG2b WB Purified 50µg 612608
WB Purified 150µg 612609
HAP1 Ms,Rat 1 MouseIgG1 IF,WB Purified 50µg 611302
HDJ-2 Hu 30 MouseIgG1 WB Purified 150µg 611873
HIP1 Hu,Ms,Rat 13 MouseIgG1 WB Purified 50µg 612314
WB Purified 150µg 612315
Chick,Dog,Hu,Ms,Rat 34 MouseIgG2a WB Purified 50µg 612316
Hip1R Ms 44 MouseIgG1 WB Purified 50µg 612118
MCM Dog,Hu,Ms,Rat 24 MouseIgG1 WB Purified 150µg 611772
150µg 611773
MnSOD Dog,Hu,Ms,Rat 19 MouseIgG1 WB Purified 50µg 611580
150µg 611581
Neuronal Pentraxin Hu,Rat 22 MouseIgG1 IF,IP,WB Purified 50µg 610369
IF,IP,WB Purified 150µg 610370
Nicastrin Chick,Dog,Hu,Ms,Rat 35 MouseIgG2a WB Purified 50µg 612290
OPA1 Hu 18 MouseIgG1 IF,WB Purified 50µg 612606
IF,WB Purified 150µg 612607
PEX1 Chick,Dog,Hu,Ms,Rat 1 MouseIgG1 WB Purified 50µg 611718
WB Purified 150µg 611719
PEX19 Hu 32 MouseIgG1 IF,WB Purified 50µg 611598
Neurotransmitter Receptor Related (continued)
description react clone isotype apps format size cat.no.
Tyrosine Hydroxylase Ms,Rat 45 MouseIgG1 WB Purified 50µg 612300
Rat TOHA1 MouseIgG1 EM,IHC(Fr) Purified 100µg 556311
VAChT Ms,Rat null Goat IHC(Fr) Serum 50µL 556337
bdbiosciences.com50 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Functional Assays and Discovery Reagents (continued)
Pathology (continued)
description react clone isotype apps format size cat.no.
SIP1 Dros 4 MouseIgG1 IF,IHC,WB Purified 50µg 612655
Hu 4 MouseIgG1 IF,WB Purified 50µg 611256
SMN Hu 8 MouseIgG1 IF,IHC,WB FITC 50µg 612586
IF,IHC,WB FITC 150µg 612587
Dog,Hu,Ms,Rat 8 MouseIgG1 IF,IHC,WB Purified 50µg 610646
IF,IHC,WB Purified 150µg 610647
TCBP-49 Ms,Rat 30 MouseIgG2a WB Purified 50µg 612178
WB Purified 150µg 612179
UBE3A Hu,Ms 13 MouseIgG1 IF,WB Purified 50µg 611416
Synaptic Vesicles
description react clone isotype apps format size cat.no.
a-Methylacyl-CoA Racemase (AMACR) Hu,Ms,Rat 15 MouseIgG1 WB Purified 50µg 612082
WB Purified 150µg 612083
Adaptin a Dog,Hu,Ms,Rat 8 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610501
IF,IHC,IP,WB Purified 150µg 610502
Adaptin b Chick,Dog,Frog,Hu,Ms,Rat 74 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610381
IF,IHC,IP,WB Purified 150µg 610382
Adaptin d Hu 18 MouseIgG1 IF,WB Purified 50µg 611328
IF,WB Purified 150µg 611329
Adaptin e Hu null IgG1 IF,WB Purified 50µg 612018
IF,WB Purified 150µg 612019
Adaptin g Dog,Hu,Ms,Rat 88 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610385
IF,IHC,IP,WB Purified 150µg 610386
AF6 Hu,Ms,Rat 35 MouseIgG2a IF,WB Purified 50µg 610732
150µg 610733
a-/b -SNAP Chick,Hu,Ms,Rat 32 MouseIgG2a IF,WB Purified 50µg 611898
Amphiphysin Ms,Rat 15 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610714
AP180 Hu,Ms,Rat 34 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610469
IF,IHC,IP,WB Purified 150µg 610470
ARF3 Chick,Dog,Frog,Hu,Ms,Rat 41 MouseIgG1 IP,IF,WB Purified 50µg 610784
b-NAP Ms,Rat 18 MouseIgG1 IF,WB Purified 50µg 610892
IF,WB Purified 150µg 610893
BRAMP2 Ms,Rat 54 MouseIgG1 IF,IP,WB Purified 50µg 610856
IF,IP,WB Purified 150µg 610857
CDCrel-1 Hu 49 MouseIgG1 IF,WB Purified 50µg 612386
Cellugyrin Ms,Rat 24 MouseIgG1 IF,WB Purified 50µg 611128
Nerves transmit signals from one to another at contact points called
synapses via a process called synaptic transmission. The two types of
synaptic transmission are electrical and chemical. Electrical transmis-
sion occurs at gap junctions. Chemical synaptic transmission occurs
at synaptic vesicles via neurotransmitters released at the presynaptic
neuron that diffuse across the synaptic cleft and are detected by
receptors on the postsynaptic cell. The postsynaptic cell can be a
neuron, a muscle, or a secretory or gland cell.
Synaptic vesicles are organelles situated at the distal terminus of the
presynaptic neuron. In the typical sequence of events, the action
potential arrives at the axon terminal and triggers influx of calcium
ions that signal exocytosis, the fusion of synaptic vesicles with the
cell membrane. SNARE complex proteins then regulate membrane
fusion. SNARE complex consists of the SNAPs (soluble NSF attach-
ment proteins), NSF (N-ethyl-maleimide-sensitive factor) along with
the receptor proteins (SNAREs such as synaptobrevin, synapto-
tagmin, syntaxin, and SNAP-25). Several Rab proteins expressed in
neuronal cells and are associated with synaptic vesicles.
Synaptic Vesicles and Post-Synaptic Proteins
bdbiosciences.com51Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Synaptic Vesicles (continued)
description react clone isotype apps format size cat.no.
Chromogranin A Rat 5 MouseIgG1 IF,WB Purified 50µg 611844
Chromogranin B Hu,Ms,Rat 32 MouseIgG1 IF,WB Purified 50µg 611646
IF,WB Purified 150µg 611647
Clathrin Heavy Chain Chick,Dog,Hu,Ms,Rat 23 MouseIgG1 IF FITC 50µg 611784
IF,IHC,IP,WB Purified 50µg 610499
IF,IHC,IP,WB Purified 150µg 610500
Complexin 2 Dog,Rat 48 MouseIgG1 IF,IP,WB Purified 50µg 610728
IF,IP,WB Purified 150µg 610729
CSP Rat 16 MouseIgG1 IF,WB Purified 50µg 612662
IF,WB Purified 150µg 612663
DLP1 Dog,Hu,Ms,Rat 8 MouseIgG1 IF,WB Purified 50µg 611112
150µg 611113
Dynamin Chick,Dog,Hu,Ms,Rat 41 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610245
IF,IHC,IP,WB Purified 150µg 610246
Dynamin II Hu,Ms,Rat 27 MouseIgG2a IF,WB Purified 50µg 610263
150µg 610264
GS15 Bov,Dog,Frog,Hu,Ms,Rat 19 MouseIgG1 IF,WB Purified 50µg 610960
IF,WB Purified 150µg 610961
GS27 Dog,Hu 25 MouseIgG1 IF,WB Purified 50µg 611034
IF,WB Purified 150µg 611035
GS28 Hu,Ms,Rat 1 MouseIgG2a IF,WB Purified 50µg 611184
IF,WB Purified 150µg 611185
Mint1 Ms,Rab,Rat 23 MouseIgG1 IF,WB Purified 50µg 611028
IF,WB Purified 150µg 611029
Mint2 Ms,Rat 18 MouseIgG1 IF,WB Purified 50µg 611032
Mint3 Hu,Ms,Rat 32 MouseIgG1 IF,WB Purified 50µg 611380
Munc-18 Rat 31 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610336
IF,IHC,IP,WB Purified 150µg 610337
Neurexin I Dog,Hu,Ms,Rat 17 MouseIgG1 IF,WB Purified 50µg 611882
NSF Chick,Dog,Hu,Ms,Rat 7 MouseIgG2b IF,WB Purified 50µg 612272
IF,WB Purified 150µg 612273
p47A Dog,Hu,Ms,Rat 26 MouseIgG1 IF,WB Purified 50µg 610890
IF,WB Purified 150µg 610891
18 MouseIgG1 IF,WB Purified 50µg 610900
panMunc13 Rat 32 MouseIgG1 IF,WB Purified 50µg 610998
IF,WB Purified 150µg 610999
ProSAP1 Rat 2 MouseIgG1 WB Purified 50µg 612274
Rab11 Chick,Dog,Hu,Ms,Rat 47 MouseIgG2a IF,IHC,WB Purified 50µg 610656
150µg 610657
Rab3 Dog,Rat 9 MouseIgG2a IF,IHC,IP,WB Purified 50µg 610379
Rab5 Hu 1 MouseIgG2a IF,IHC,WB Purified 50µg 610724
150µg 610725
Rim Ms,Rat 26 MouseIgG1 IF,WB Purified 50µg 610906
IF,WB Purified 150µg 610907
Sec8 Chick,Dog,Hu,Ms,Rat 14 MouseIgG2b IF,IP,WB Purified 50µg 610658
150µg 610659
SNAP-25 Ms,Rat 20 MouseIgG1 IF,IP,WB Purified 50µg 610366
IF,IP,WB Purified 150µg 610367
bdbiosciences.com52 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Functional Assays and Discovery Reagents (continued)
Post-Synaptic Proteins
description react clone isotype apps format size cat.no.
Arc Rat 40 MouseIgG1 WB Purified 50µg 612602
WB Purified 150µg 612603
Gephyrin Chick,Hu,Ms,Rat 45 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610584
IF,IHC,IP,WB Purified 150µg 610585
Dros 45 MouseIgG1 IF,IHC,IP,WB Purified 50µg 612632
GRIP Rat 32 MouseIgG1 IF,WB Purified 50µg 611318
IF,WB Purified 150µg 611319
MUPP1 Ms,Rat 43 MouseIgG1 IF,WB Purified 50µg 611558
IF,WB Purified 150µg 611559
Neurabin Ms,Rat 52 MouseIgG1 IF,WB Purified 50µg 611088
Dros 52 MouseIgG1 IF,WB Purified 50µg 612639
Neurabin II Rat 17 MouseIgG1 WB Purified 50µg 612166
PSD-95 Ms,Rat 16 MouseIgG1 IF,IP,WB Purified 50µg 610495
IF,IP,WB Purified 150µg 610496
SCAMP1 Hu 22 MouseIgG1 WB Purified 150µg 612087
Vesl-1L Rat 8 MouseIgG1 IF,WB Purified 50µg 611174
Synaptic Vesicles (continued)
description react clone isotype apps format size cat.no.
Synapsin I Ms,Rat 8 MouseIgG1 IF,WB Purified 50µg 611392
IF,WB Purified 150µg 611393
Synapsin IIa Hu,Rat 1 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610666
Synaptogyrin Rat 6 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610597
Synaptophysin Ms,Rat 2 MouseIgG1 WB Purified 50µg 611880
Synaptotagmin Hu,Rat 41 MouseIgG1 IF,IP,WB Purified 50µg 610433
IF,IP,WB Purified 150µg 610434
Synaptotagmin V Ms,Rat 46 MouseIgG1 WB Purified 50µg 612284
Syncollin Rat 17 MouseIgG2b WB Purified 50µg 612468
Syndapin I Ms,Rat 32 MouseIgG1 WB Purified 50µg 611810
WB Purified 150µg 611811
Syntaphilin Ms,Rat 30 MouseIgG1 WB Purified 50µg 612324
Syntaxin 11 Hu,Ms 32 MouseIgG1 IF,WB Purified 50µg 611282
IF,WB Purified 150µg 611283
Syntaxin 4 Dog,Hu,Ms,Rat 49 MouseIgG1 IF,IP,WB Purified 50µg 610439
IF,IP,WB Purified 150µg 610440
Syntaxin 6 Chick,Dog,Frog,Hu,Ms,Rat 30 MouseIgG1 IF,IP,WB Purified 50µg 610635
IF,IP,WB Purified 150µg 610636
Syntaxin 8 Dog,Hu,Ms,Rat 48 MouseIgG2a IF,WB Purified 50µg 611352
IF,WB Purified 150µg 611353
Tomosyn Hu,Ms,Rat 15 MouseIgG1 IF,WB Purified 50µg 611296
IF,WB Purified 150µg 611297
bdbiosciences.com53Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
For Probenecid Sensitive Applications
description size cat.no.
BD PBX Calcium Assay Kit 10plates 640175
100plates 640177
For Standard Applications
description size cat.no.
BD Calcium Assay Kit 10plates 640176
100plates 640178
Assay Kits and Reagents for Bioimaging and Flow Cytometry
BD introduces two new calcium assay kits for detecting calcium flux
in living cells. The proprietary formulations combine a non-fluores-
cent calcium indicator that becomes activated inside the cell, along
with a signal enhancer to provide an easy-to-use, no-wash assay
format with increased signal to background. In most cases, the dye
kits perform better than kits currently available on the market with
no known interference to ligands (small molecules, peptide/proteins,
and lipids). Also, the “addition artifact” commonly seen with some
kits is significantly reduced with these new dye formulations.
• No-wash homogeneous assay format
• Higher signal over background compared to competitive kits
• PBX kit permits assays with reduced or no probenecid
• Ideal for all fluorescence-based calcium flux assays
The kits have been successfully used with a variety of cell lines
including CHO, HEK293, Jurkat, Hela, and U2OS.
Kits are available in two sizes providing sufficient material for either
10 plates (96, 384, or 1536 wells) or 100 plates. Additionally, for
higher volume applications, custom packaged materials can also be
provided. Please contact customer support for more details.
Calcium Flux Assays
bdbiosciences.com54 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Functional Assays and Discovery Reagents (continued)
BD Biosciences has always provided the most reliable and complete
multiplex bead array system. We have taken the performance you
trust and the ease of use you rely on and have made them even
more flexible. In addition to your need for a flexible assay system,
we also recognize the value of your precious samples and the
importance of data reproducibility, so we’ve engineered the new
BD™ Cytometric Bead Array (CBA) Flex Set system to maximize
both. Further, BD CBA Flex Sets can be used with any dual-laser
flow cytometer, or you can increase throughput by automating your
acquisition and analysis on a BD FACSArray™ bioanalyzer.
With the BD CBA Flex Set System You Can:
• Configure your multiplex assay on demand
• Get multiple results from a single, small-volume sample
• Achieve quantitative results in less time
• Use with any dual-laser flow cytometer (non stream-in-air)
• Automate sample acquisition and increase throughput with the
BD™ HTS option coupled with BD™ flow cytometers or the
plate-based BD FACSArray bioanalyzer
The new BD CBA Flex Set system is an open and configurable system
designed to be the easiest method of creating multiplex assays.
With our proprietary
conjugation chemistry, pair
optimization strategies,
and direct PE detection
reagents, you can be
assured consistent and
superior assay performance
in complex biological
samples. Each antibody
pair we develop is
evaluated for dynamic
range, sensitivity, and
parallel titration to native
biological samples. We have also specifically formulated the assay
diluent and wash buffers for each assay type to reduce potentially
detrimental effects on assay performance when using serum and
plasma samples. By avoiding the streptavidin-biotin-PE detection
method employed by other assays, our direct PE detection reagents
minimize the risk of increased background often caused by
endogenous biotin in serum and lysate samples. Thus, BD CBA Flex
Sets provide a reliable and flexible method for quantitative detection
of multiple analytes in a single serum, plasma, tissue culture
supernatant, or cell lysate sample.
102 10 103 4 105
Red-A
2
3
4
10
10
10
10
5
NIR
-A
BD Cytometric Bead Array Flex Set Reagents
description react bead size cat.no.
Btk (Y551) Hu D5 100tests 558236
eNOS (S1177) Hu C7 100tests 558239
ERK1/2 (T202/Y204) Hu,Ms,Rat C4 100tests 558234
Itk (Y511) Hu,Ms C6 100tests 558230
JNK1/2 (T183/Y185) Hu,Ms,Rat B5 100tests 558235
p38 (T180/Y182) Hu,Ms,Rat B6 100tests 558233
PLC-g (Y783) Hu,Ms,Rat B7 100tests 558228
RSK (T573) Hu,Ms,Rat D7 100tests 558240
Stat1 (Y701) Hu,Ms C5 100tests 558222
Syk (Y352) Hu,Ms,Rat B9 100tests 558237
ZAP-70 (Y319) Hu,Ms B8 100tests 558229
Stat1 Hu,Ms,Rat D4 100tests 558227
Syk Hu,Ms,Rat B9 100tests 558239
ZAP-70 Hu,Ms,Rat B8 100tests 558232
Cell Signaling Master Buffer Kit 100tests 558223
500tests 558224
BD Cytometric Bead Array Flex Set Reagents
Visit bdbiosciences .com/flexset for complete product listings.
bdbiosciences.com55Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
High-Content Bioimaging Certified Antibodies
description clone apps format size cat.no.
53BP1 19 IF,WB Purified 50µg 612522
IF,WB Purified 150µg 612523
b-Tubulin 5H1 ICC,WB Purified 100µg 556321
Actin C4 IF,WB Purified 50µg 612656
IF,WB Purified 150µg 612657
ATP Synthase b 10 IF,WB Purified 50µg 612518
IF,WB Purified 150µg 612519
Bcl-x 44 IF,IHC,IP,WB Purified 50µg 610746
IF,IHC,IP,WB Purified 150µg 610747
Bid 7 IF,WB Purified 50µg 611528
IF,WB Purified 150µg 611529
Bub3 31 IF,WB Purified 50µg 611730
IF,WB Purified 150µg 611731
c-IAP-1 B75-1 IP,WB Purified 100µg 556533
c-Jun (pS63) 2 IHC(F),WB Purified 100µg 558036
Cab45 30 IF,WB Purified 50µg 611064
IF,WB Purified 150µg 611065
Calnexin 37 IF,IHC,IP,WB Purified 50µg 610523
IF,IHC,IP,WB Purified 150µg 610524
Calretinin 34 IF,WB Purified 50µg 610908
IF,WB Purified 150µg 610909
Caspase-2 G310-1248 WB Purified 50µg 551093
WB Purified 150µg 551094
Caspase-3 19 IF,IHC,IP,WB Purified 50µg 610322
IF,IHC,IP,WB Purified 150µg 610323
Caspase-3, Active Form C92-605 FCM,IHC(Fr),IP Purified 25µg 559565
Caspase-6 B93-4 WB Purified 100µg 556581
Caspase-7 8-1-47 IP,WB Purified 50µg 551236
IP,WB Purified 150µg 551237
Caspase-8 Polyclonal WB Serum 100µL 551234
Caspase-14 32 IF,WB Purified 50µg 611510
IF,WB Purified 150µg 611511
Caveolin 1 C060 FCM,IF,IHC,IP,WB Purified 50µg 610057
FCM,IF,IHC,IP,WB Purified 150µg 610058
Cdc25B 23 IF,IHC,IP,WB Purified 50µg 610527
IF,IHC,IP,WB Purified 150µg 610528
Cdc25C C2-2 WB Purified 100µg 556576
CDC5L 21 IF,WB Purified 50µg 612362
BD Biosciences is now offering a rapidly growing collection of
antibody reagents that have been tested and certified for research
applications in automated bioimaging (high-content screening, HCS)
and fluorescence microscopy.
These Bioimaging Certified reagents are optimal for use in
high-content applications when reproducibility is essential.
BD Biosciences bioimaging reagents are certified for optimal
performance based on an internal test program. For BD to certify
these reagents, each antibody must pass a number of tests that
focus on high content applications.
Theses tests include:
• Exceeding a signal intensity threshold over background in
commonly used cell lines
• Working with widely used fixation and permeabilization
methods
• Localizing to the appropriate region within a cell
• Showing an appropriate response (ie, redistribution) following
stimulation
Visit bdbiosciences .com/bcr for the latest product releases.
bdbiosciences.com56 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Functional Assays and Discovery Reagents (continued)
High-Content Bioimaging Certified Antibodies (continued)
description clone apps format size cat.no.
Cdk2 55 IF,IHC,IP,WB Purified 50µg 610145
IF,IHC,IP,WB Purified 150µg 610146
Cdk4 DCS-35 IF,IP,WB Purified 100µg 559693
Cdk7 MO-1 IF,IHC(Fr),IP,WB Purified 100µg 556345
Cdk8 F9-1075 WB Purified 50µg 552056
WB Purified 150µg 552053
CGBP 35 IF,WB Purified 50µg 612334
IF,WB Purified 150µg 612335
Chk2 19 IF,WB Purified 50µg 611570
IF,WB Purified 150µg 611571
Crk 22 IF,IHC,IP,WB Purified 50µg 610035
IF,IHC,IP,WB Purified 150µg 610036
Cyclin A 25 IF,WB Purified 50µg 611268
IF,WB Purified 150µg 611269
Cyclin B1 GNS-11 FCM,IF,IP,WB Purified 100µg 554178
FCM,IF,IP,WB Purified 250µg 554179
Cyclin B1 (a.a 1-21) GNS-1 FCM,IF,IHC(F), Purified 100µg 554176 IHC(Fr),IP,WB
FCM,IF,IHC(F), Purified 250µg 554177 IHC(Fr),IP,WB
Cyclin E HE12 FCM,WB Purified 50µg 551159
FCM,WB Purified 150µg 551160
Cytochrome c 6H2.B4 IP Purified 100µg 556432
D4-GDI Polyclonal WB Serum 100µL 556498
eIF-4E 87 IF,IP,WB Purified 50µg 610269
IF,IP,WB Purified 150µg 610270
IF,IP FITC 50µg 611624
IF,IP FITC 150µg 611625
eIF-5a 26 WB Purified 50µg 611976
WB Purified 150µg 611977
eNOS (pS1177), Phospho-Specific 19 FCM,WB Purified 50µg 612392
FCM,WB Purified 150µg 612393
eNOS (pS633) 37 WB Purified 50µg 612664
WB Purified 150µg 612665
eNOS/NOS Type III 3 IF,IHC,IP,WB Purified 50µg 610296
IF,IHC,IP,WB Purified 150µg 610297
33 IF,IP,WB Purified 50µg 610427
IF,IP,WB Purified 150µg 610428
ERK (pan) 16 IF,IHC,IP,WB Purified 50µg 610123
IF,IHC,IP,WB Purified 150µg 610124
ERK1/2 (pT202/pY204) 20A WB Purified 50µg 612358
WB Purified 150µg 612359
ERK3 30 IF,IHC,IP,WB Purified 50µg 610105
IF,IHC,IP,WB Purified 150µg 610106
Ezrin 18 IF,IHC,IP,WB Purified 50µg 610602
IF,IHC,IP,WB Purified 150µg 610603
FADD 1 IF,IP,WB Purified 50µg 610399
IF,IP,WB Purified 150µg 610400
bdbiosciences.com57Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
High-Content Bioimaging Certified Antibodies (continued)
description clone apps format size cat.no.
FAK 77/FAK IF,IHC,IP,WB Purified 50µg 610087
IF,IHC,IP,WB Purified 150µg 610088
FAK (pY397) 14 IF,WB Purified 50µg 611722
IF,WB Purified 150µg 611723
18 IF,WB Purified 50µg 611806
IF,WB Purified 150µg 611807
Fatty Acid Synthase 23 IF,WB Purified 50µg 610962
IF,WB Purified 150µg 610963
FEN-1 21 IF,WB Purified 50µg 611294
IF,WB Purified 150µg 611295
FIN13 7 IF,WB Purified 50µg 610910
IF,WB Purified 150µg 610911
Fnk 53 IF,WB Purified 50µg 611178
IF,WB Purified 150µg 611179
Fractin Polyclonal IHC,WB Purified 100µL 551527
GRB2 81 IF,IHC,IP,WB Purified 50µg 610111
IF,IHC,IP,WB Purified 150µg 610112
GSK-3b 7 IF,IHC,IP,WB Purified 50µg 610201
IF,IHC,IP,WB Purified 150µg 610202
GSK-3b (pY216) 13a WB Purified 50µg 612312
WB Purified 150µg 612313
Hsp60 24 IF,WB Purified 50µg 611562
IF FITC 50µg 611958
IF,WB Purified 150µg 611563
IKKa B78-1 IP,WB Purified 100µg 556532
IRAK 7 IF,IP,WB Purified 50µg 610754
IF,IP,WB Purified 150µg 610755
KAP 39 IF,IHC,IP,WB Purified 50µg 610334
Laminin B2 22 IF,IP,WB Purified 50µg 610722
MCM6 1 IF,WB Purified 50µg 611622
IF,WB Purified 150µg 611623
MDM2 SMP14 IF,IHC(F),IHC(Fr), Purified 100µg 556353 IP,WB
MEK1 25 IF,IHC,IP,WB Purified 50µg 610121
IF,IHC,IP,WB Purified 150µg 610122
Metablastin 53 IF,WB Purified 50µg 611146
IF,WB Purified 150µg 611147
NF-kB p65 20 IF,WB Purified 50µg 610868
IF,WB Purified 150µg 610869
p19Skp1 52 IF,IHC,IP,WB Purified 50µg 610530
p23 16 WB Purified 50µg 612320
WB Purified 150µg 612321
p27Kip1 G173-524 IP,WB Purified 100µg 554069
p32 20 IF,WB Purified 50µg 612396
IF,WB Purified 150µg 612397
p38 MAPK (pT180/pY182) 36 IF,WB Purified 50µg 612288
IF,WB Purified 150µg 612289
p47A 26 IF,WB Purified 50µg 610890
IF,WB Purified 150µg 610891
bdbiosciences.com58 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Functional Assays and Discovery Reagents (continued)
High-Content Bioimaging Certified Antibodies (continued)
description clone apps format size cat.no.
p53 DO-1 IHC(F),IHC(Fr), Purified 100µg 554293 IP,WB
p54nrb 3 IF,WB Purified 50µg 611278
IF,WB Purified 150µg 611279
p55Cdc 41 IF,WB Purified 50µg 611242
IF,WB Purified 150µg 611243
PARP 4C10-5 Blot,FCM,IP,WB Purified 100µg 556494
PARP, Cleaved Form F21-852 FCM,IP,WB Purified 50µg 552596
FCM,IP,WB Purified 150µg 552597
Paxillin 177 IF,IP,WB Purified 50µg 610568
IF,IP,WB Purified 150µg 610569
PCNA PC10 IC/FCM,IHC(F), Purified 100µg 555566 IHC(Fr)
IC/FCM Biotin 100µg 555567
Phospholipase Cg1 10 IF,IHC,IP,WB Purified 50µg 610027
IF,IHC,IP,WB Purified 150µg 610028
PI3-Kinase 4 IF,IHC,IP,WB Purified 50µg 610045
IF,IHC,IP,WB Purified 150µg 610046
PI3-Kinase C2b 22 IF,WB Purified 50µg 611342
IF,WB Purified 150µg 611343
PI4-Kinase b 7 IF,WB Purified 50µg 611816
IF,WB Purified 150µg 611817
PKARI 18 IF,IHC,IP,WB Purified 50µg 610165
IF,IHC,IP,WB Purified 150µg 610166
IF FITC 50µg 610167
IF FITC 150µg 610168
PKARIIa 40 IF,IP,WB Purified 50µg 612242
IF,IP,WB Purified 150µg 612243
Plakophilin 2 28 IF,IP,WB Purified 50µg 610788
IF,IP,WB Purified 150µg 610789
PNUTS 47 IF,WB Purified 50µg 611060
IF,WB Purified 150µg 611061
Poly ADP-Ribose 10H IF Purified 50µg 550781
Psme3 47 IF,WB Purified 50µg 611180
IF,WB Purified 150µg 611181
PTRF 4 IF,WB Purified 50µg 611258
IF,WB Purified 150µg 611259
Rac1 102 IF FITC 50µg 610652
IF FITC 150µg 610653
RACK1 20 IF,IHC,IP,WB Purified 50µg 610177
IF,IHC,IP,WB Purified 150µg 610178
Ran 20 IF,IHC,IP,WB Purified 50µg 610340
IF,IHC,IP,WB Purified 150µg 610341
RanBP3 52 IF,WB Purified 50µg 612050
IF,WB Purified 150µg 612051
RAP30 15 IF,WB Purified 50µg 612176
IF,WB Purified 150µg 612177
Rb (a.a. 300-380) G3-349 IP,WB Purified 100µg 554140
Rb (a.a. 332-344) G3-245 FCM,IF, Purified 100µg 554136 IC/FCMBlock,IHC(F), IHC(Fr),IP,GS,WB
bdbiosciences.com59Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
High-Content Bioimaging Certified Antibodies (continued)
description clone apps format size cat.no.
S100P 16 IF,IHC,IP,WB Purified 50µg 610306
IF,IHC,IP,WB Purified 150µg 610307
Sec31A 32 IF,WB Purified 50µg 612350
IF,WB Purified 150µg 612351
SII 7 IF,WB Purified 50µg 611204
IF,WB Purified 150µg 611205
Smac/DIABLO 7 IF,WB Purified 50µg 612244
IF,WB Purified 150µg 612245
Sos1 25 IF,IHC,IP,WB Purified 50µg 610095
IF,IHC,IP,WB Purified 150µg 610096
SRC-1 8 IF,WB Purified 50µg 612378
Stat1 42 IF,IHC,IP,WB Purified 50µg 610185
IF,IHC,IP,WB Purified 150µg 610186
Stat1 (pY701) 14 FCM,IF,IP,WB Purified 50µg 612132
FCM,IF,IP,WB Purified 150µg 612133
4a FCM,IF,IP,WB Purified 50µg 612232
FCM,IF,IP,WB Purified 150µg 612233
Stat3 (pS727) 49 IF,WB Purified 50µg 612542
IF,WB Purified 150µg 612543
Stat3 (pY705) 4 WB Purified 50µg 612356
WB Purified 150µg 612357
Stat6 23 IF,WB Purified 50µg 611290
IF,WB Purified 150µg 611291
Stat6 (pY641) 18 IF,WB Purified 50µg 611566
IF,WB Purified 150µg 611567
TOK-1 13 WB Purified 50µg 612326
Topo IIa 31 IF,WB Purified 50µg 611326
IF,WB Purified 150µg 611327
Topo IIb 40 IF,WB Purified 50µg 611492
IF,WB Purified 150µg 611493
Visit bdbiosciences .com/bcr for our latest product releases.
bdbiosciences.com60 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Functional Assays and Discovery Reagents (continued)
Screening and Development of Antibodies for use in Cell-Based Assays using Immunofluorescence Microscopy
IntroductionHigh content imaging is rapidly becoming a mainstay in pharmaceutical and life science research laboratories.1 The power of imaging resides in its ability to measure not only fluorescence intensity changes within cells, but also molecular rearrangements and morphological features. In recent years, the availability of a new generation of automated imaging platforms has created a unique opportunity to develop novel cell-based assays.2-4 High content microscopy has also been advanced by the abundant use of antibodies in immunofluorescence applications. These antibody reagents have enabled high resolution localization of important protein targets in individual cells. For example, antibodies can be used to detect specific stages in apoptosis, in the activation of signaling cascades, and in the cell cycle. However, many of the antibodies currently available have not been specifically tested for use in automated imaging applications. In addition, the promise of high content screening relies on the multiplexing of cellular information; the ability to do this however, is often limited when using standard unlabeled primary antibodies in combination with labeled secondary antibodies due to species cross-reactivity.
To facilitate the use of antibodies for imaging applications, BD Biosciences embarked on a screening process using automated imaging to evaluate a large library of monoclonal antibodies. The library of antibodies included specificities that recognize proteins involved in apoptosis, cancer, cell cycle, cell signaling, and neurobiology. Antibodies were tested using cell lines and methods that are relevant in automated imaging. Specific criteria regarding signal to noise, sub-cellular localization, and other important parameters were used to qualify reagents. Two distinct Bioimaging Certified Reagent product lines have resulted from this antibody screen. The first is a collection of more than 200 unlabeled primary antibodies that have been shown to have general utility in bioimaging applications. A subset of these reagents has been developed into conjugated primary antibodies in order to truly enable high content screening using antibody multiplexing. These reagents, combined with an automated imaging platform such the BD Pathway bioimager 415, 435, or 855 can enable rapid development of novel cell-based assays.
Methods for Screening of Purified AntibodiesAntibodies from the BD Biosciences library were screened for utility in bioimaging applications. The antibodies recognized proteins in different areas of biology that included apoptosis, cancer, cell cycle, cell signaling, and neurobiology. For ubiquitously expressed proteins, the antibodies were screened on three different cell lines routinely used in high content imaging applications. HeLa (human cervical carcinoma), A549 (human lung carcinoma), or U-2 OS (human osteosarcoma) cells were seeded at approximately 10,000 cells/well in 96-well clear bottom tissue culture plates (Cat. No. 353219) optimized for automated imaging applications. Eighteen hours later, cells were either treated with an appropriate compound and then fixed and permeabilized or, if not treated, immediately fixed and permeabilized.
Fixation
1. Remove culture media from wells.
2. Add 100 μL freshly prepared 3.7% formaldehyde diluted in 1x phosphate buffered saline (PBS) (Sigma F-1268) and pre-warmed to 37°C to each well.
3. Incubate at room temperature for 10 minutes.
4. Remove the formaldehyde solution and drain the residual liquid by inverting the plate briefly onto absorbent paper.
5. Wash by adding 100 μL per well of 1x PBS.
6. Proceed to either the Triton® X-100 or Methanol permeabiliza-tion steps described below.
Triton® X-100 Permeabilization
1. Remove 1x PBS and add 100 μL per well of a 0.1% Triton® X-100 solution diluted in 1x PBS.
2. Incubate at room temperature for 5 minutes.
3. Remove the 0.1% Triton® X-100 solution and drain the residual liquid.
4. Wash twice with 1x PBS.
5. Proceed to the Blocking step.
Methanol Permeabilization
1. Remove 1x PBS and add 100 μL per well of a -20°C 90% meth-anol solution diluted in water.
2. Incubate at room temperature for 5 minutes.
3. Remove the 90% methanol solution and drain the residual liquid.
4. Wash twice with 1x PBS.
5. Proceed to the Blocking step.
bdbiosciences.com61Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Technical Spotlight and Protocols
bdbiosciences.com62 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Blocking
1. Remove 1x PBS and add 100 μL per well of 1x PBS supple-mented with 3% fetal bovine serum (FBS blocking solution).
2. Incubate at room temperature for 30 minutes.
3 Proceed to the Primary Antibody Staining step.
Primary Antibody Staining
1. Remove the 3% FBS blocking solution and add 50 μL per well of primary antibody diluted to the appropriate concentration in blocking solution.
2. Incubate at room temperature for 1 hour.
3. Remove primary antibody solution.
4. Wash three times with the blocking solution.
5. Proceed with Secondary Antibody Staining.
Secondary Antibody Staining
1. Remove the 3% FBS blocking solution and add 50 μL per well of diluted secondary antibody (diluted in blocking solution). Several different second step reagents were used, including FITC goat anti-mouse IgG (Cat. No. 554001), Alexa Fluor® 555 goat anti-mouse IgG (Invitrogen), and Alexa Fluor® 488 goat anti-mouse IgG (Invitrogen). All were used at 0.25μg/well.
2. Incubate at room temperature for 1 hour in the dark.
3. Remove secondary antibody solution.
4. Wash three times with the blocking solution.
5. Add 100 μL per well of 1x PBS containing 2 μg/mL Hoechst 33342 (Sigma).
6. Cover the plate and let stand protected from light for 15 to 30 minutes.
7. Plates are now ready for imaging.
Screening of Directly Conjugated Antibodies
Essentially the same procedure as outlined above was followed, with the omission of the secondary antibody step, as the primary antibodies were directly conjugated to Alexa Fluor® dyes. After incubation with a conjugated antibody, cells were washed as described, Hoechst dye was added, and the samples were imaged.
Imaging and Analysis
Cells were imaged on a BD Pathway 855 bioimager using a 20x (0.75 NA) objective. Image analysis was done using the built in Attovision (version 1.5) software tools. Numerical data was imported into the Microsoft® Excel® based BD™ Image Data Explorer software, where cell populations were analyzed for nuclear and/or cytoplasmic fluorescence intensity.
Technical Spotlight and Protocols (continued)
Erk1/2 (pT202/pY204) During Differentiation of P19 Cells
100 101 102 103 104
Red-A
100 101 102 103 104
Red-A
100 101 102 103 104
Red-A
P19 P19+DMSO P19+Retinoic Acid
Figure 1 . Undifferentiated (left panel), DMSO differentiated (middle panel), and retinoic acid differentiated (right panel) P19 cells, were serum starved for 2 hours (orange), then treated with either PMA (green) or NGF (blue). Cells without serum starvation (red) were also included for comparison.
Erk1/2 (pT202/pY204) During Differentiation of P19 Cells
Phospho-Protein Profiling in Stem Cells
Characterization of the spatial and temporal function, or proteomic
profiling, of each component of the cellular proteome is one of the
major scientific challenges faced today. This proteomic profiling is of
critical importance in understanding how pluripotent cells develop
into an adult and complete organism, or replenish and support
existing organs. Those pluripotent cells, or stem cells, undergo a
timely coordinated gene expression inducing their differentiation.
Understanding the signaling mechanisms orchestrating those
dynamic intracellular changes is key to comprehending the biology
of stem cells. Reversible protein phosphorylation, controlled by
kinases and phosphatases, has been recognized as a universal
post-translational modification that rapidly adjusts the activity of a
given protein target. Yet despite its central role in cell biology, little
is known about the role of protein phosphorylation in stem cells and
their differentiation. Early studies demonstrated the importance of
phospho-STAT3,79 phospho-Erk, and the Src kinase in maintaining
the pluripotency of stem cells.80 Furthermore, FAK (focal-adhesion
kinase), is required for cardiogenesis of mouse embryonic stem
cells.81 Most methodologies used to monitor protein phosphorylation
in differentiating stem cells lack the ability to visually represent the
variability of this modification of individual cells and committed cell
sub-populations, thus limiting data interpretation.
BD Phosflow technology is an emerging methodology whereby the
phosphorylation of individual proteins can be monitored by flow
cytometry using highly specific antibodies as probes. The ability of
flow cytometry to identify unique cell populations opens the
possibilities for monitoring protein phosphorylation in rare cell
events types, such as T-cell precursors, developing cells, and
malignant stem cells.82,83
In the experiment described below, we have used mouse
pluripotent embryonic carcinoma stem cells (P19) to demonstrate
that BD Phosflow technology is a valuable tool for the analysis of
differentiation stages in stem cells. The P19 cell line is derived from
an embryonal carcinoma induced in a C3H/He strain mouse. The
pluripotent P19 cells can be induced to differentiate into neuronal
and glial cells in the presence of retinoic acid.84 Aggregates of P19
cells differentiate into cardiac and skeletal muscle in the presence of
dimethyl sulfoxide (DMSO). In the presence of both retinoic acid and
DMSO, cells develop as if exposed to retinoic acid only.
In Figure 1, undifferentiated, DMSO differentiated, and retinoic
acid differentiated P19 cells were serum starved for two hours. After
stimulation with either phorbol myristate acetate (PMA) or nerve
growth factor (NGF), the cells were fixed, permeabilized, stained
with Alexa Fluor® 647 conjugated anti-phospho ERK1/2
(Cat. No. 612593), and subsequently analyzed by flow cytometry.
The data shown indicates that only retinoic acid differentiated cells
show a significant increase in Erk1/2 phosphorylation upon stimula-
tion with NGF. No significant effect on Erk1/2 phosphorylation could
be detected in either retinoic acid or DMSO differentiated cells after
treatment with PMA.
Technical Spotlight
bdbiosciences.com63Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
bdbiosciences.com64 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Related Immunohistochemistry Reagents
description size cat.no.
IHC Zinc Fixative 1000mL 550523
10× Zinc Fixative 500mL 552658
A ready-to-use solution, our IHC zinc fixative is an alternative to the harsh conditions of formalin fixation .
Immunohistochemistry (IHC) is a powerful technique used by
neuroscientists to identify neuron connections and to elucidate the
role of chemical messengers in relation to the architecture of the
brain. IHC with free-floating tissue sections is often impractical,
however, because it requires large amounts of antibody and
prolonged incubation periods. BD Pharmingen and
BD Transduction Laboratories antibodies, using our formalin-
free IHC Zinc Fixative (Cat. No. 550523), allow staining
to be performed on 5-μm, paraffin-embedded tissue
sections. The protocol is quick and easy to perform, the
morphologies obtained are superior to those obtained with
free-floating sections, and tissue antigenicity is retained.
In addition, our IHC antibodies are supported by a complete line of
ancillary reagents such as secondary antibodies, isotype controls, and
detection systems.
See images on the following page for phospho-specific staining of
rat brain tissues.
Visit our technical support protocol website at bdbiosciences .com
for the complete protocol on preparation and staining of
paraffin sections.
Glutamate receptor mGluR1a staining on normal rat cerebellum using our IHC Zinc Fixative: A) free-floating section and B) zinc-fixed, paraffin-embedded section. Magnification: 1× (left) and 10× (right).
A A
B B
IHC for Neuroscience Research
Technical Spotlight and Protocols (continued)
bdbiosciences.com65Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Phospho-Specific Staining of Rat Brain Tissues
ERK1 IHC images and accompanying Western blot images
ERK2 IHC image and accompanying Western blot image
JUN IHC image and accompanying Western blot image
nNos IHC image and accompanying Western blot image
Calcimeurin IHC image and accompanying Western blot image
Imaging and High Content Analysis
BD CARV II Confocal Imager
The BD CARV II confocal imager delivers high-resolution CCD
confocal imaging in an easy-to-use and cost-effective optical
package that fits on your existing microscope. High-speed multi-
point confocal scanning, combined with high quantum efficiency
CCD cameras, minimizes photobleaching and allows real-time
imaging and recording at up to 100 fps. A long life arc source
coupled to the instrument via an alignment-free light guide allows
for full spectrum (360–700 nm) confocal imaging of virtually any
fluorescent probe. Automation of internal multi-position excitation,
dichroic, and emission filter wheels permits fast multi-dimensional
imaging of up to five fluorescent probes in the same sample.
BD Pathway Bioimager
The BD Pathway bioimager is an automated, confocal, real-time,
single-cell kinetic and endpoint imaging system that has been inte-
grated into a single, compact unit. The BD Pathway bioimager was
designed to provide high-resolution, automated confocal imaging
with sophisticated imaging software to assist in developing better
cell-based assays. The system allows the user to explore biological
events without many of the restrictions of conventional microscope-
based high-content imaging systems.
The BD CARV II system provides an easy-to-use confocal upgrade to your existing microscope without sacrificing image quality or three-dimensional imaging capabilities.
The BD Pathway bioimager provides a wide range of imaging capa-bilities including kinetic, endpoint, and three-dimensional confocal imaging. The environmentally controlled imaging chamber with liquid handling provides unsurpassed live cell kinetic imaging possibilities.
Bioimaging Instrumentation
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Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
bdbiosciences.com67Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
BD FACSArray Bioanalyzer
Quantitative multi-parameter cellular analysis is conducted using
flow cytometry instruments by analyzing whole cells and proteins
separately or in concert. This technique is creating new ways to
analyze neuronal cells from small sample sizes. The BD FACSArray
bioanalyzer is the primary instrument for this application.
Unlimited Possibilities
The BD FACSArray bioanalyzer from BD Biosciences, the innovation
leader in flow cytometry instrumentation, is used for fast and sensi-
tive high-content measurements of cells and proteins in cell biology,
immunology, and proteomics.
This compact flow cytometer rapidly detects and quantifies
concentrations of secreted proteins, proteins in cell lysates,
and cell-associated proteins using small sample volumes
directly from a 96-well microtiter plate through multiplexed
BD Cytometric Bead Arrays (CBAs). Furthermore, it supports
your search for answers in cellular analysis of apoptosis,
cytokine and chemokine profiling, and phosphorylation of key
signal transduction proteins. Our bioanalyzer can also be used to
assess hybridomas, cell viability, proliferation, and immunotoxicology.
For your lab, this means unlimited possibilities for both cellular
applications and multiplex bead assays.
See page 55 or visit bdbiosciences .com/flexset for a list of
BD CBA reagents.
The BD Biosciences commitment to application development is one
of our strongest distinctions in supporting customers. Carefully opti-
mized applications and availability of over 1,000 reagents support
the BD FACSArray bioanalyzer platform.
A fully integrated system, the BD FACSArray bioanalyzer is composed
of an analyzer with an automated microtiter plate sampler, a sensi-
tive optical detection subsystem, a computer workstation, intuitive
software, and validated applications. It is easy to use and requires
minimal training, so your research shows immediate success.
BD FACSArray bioanalyzer system:• Fast microtiter plate sampler
• Two-laser system
• Sensitive fluorescence detection
• Simultaneous collection of up to six parameters per particle (four fluorescence and two scatter)
• Intuitive software
• State-of-the-art digital signal processing allows up to 15,000 events per second
• Supported by BD Biosciences reagents and applications
Application Flexibility
Cellular Analysis Using Flow Cytometry
Maximize Your Productivity
The BD FACSArray bioanalyzer features an automated microtiter
plate sampler that quickly acquires from 96-well plates. The plate
sampler is controlled by flexible and easy-to-use software for imme-
diate adoption of the bioanalyzer in your laboratory. The system
software controls the instrument and sample acquisition, and is
powerful enough to support batch analysis across multiple plates.
Its experiment wizard contains ready-to-use acquisition and analysis
templates for key applications, and lets you rapidly define and store
your own templates. One-click instrument maintenance protocols
make it easy to keep the bioanalyzer performing optimally.
BD Biosciences assays for the BD FACSArray bioanalyzer are devel-
oped with consideration for simplicity and small sample volume,
while taking advantage of the analyzer’s multiparameter detection
capability which results in many answers per sample.
System speed, novel software tools, and streamlined assay formats
accelerate your total productivity and interpretation of high-content
biological data for the answers you need.
Discover the Flexibility
The BD FACSArray bioanalyzer has an optical detection system that
is ideal for multicolor flow cytometry experiments. The optics are
capable of collecting up to four fluorescent colors from each sample,
as well as forward and side scatter light. Correlated forward and
side scatter signals are crucial for differentiating events of interest
from interfering debris. With its multiparameter capabilities, you can
resolve various populations within a single cellular or bead sample.
For multiplex bead applications, antibody-labeled fluorescent beads
capture and quantify secreted proteins or proteins in cell lysates
using BD CBA technology. With BD CBA Flex Set assays, you tailor
the bead array to your specific experiment. For cellular applications,
antibodies conjugated to fluorophores bind specifically to cell surface
or intracellular proteins. This technique can be applied to resting,
immune-challenged, or drug-exposed samples to differentiate cells
and to define their biological responses. Additionally, dyes can be
used to determine cell viability. BD Biosciences applications cover the
entire range of species including studies of human, murine, dog, and
non-human primate samples.
bdbiosciences.com68 Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.
Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
Imaging and High Content Analysis (continued)
*ClassI(1)laserproduct†Forinvitrodiagnosticuse‡Seven-andeight-colorimmunophenotypingrequirevalidationbyuser.§Forresearchuseonly.Notforuseindiagnosticortherapeuticprocedures.
The BD™ High Throughput Sampler (HTS) option available for the BD FACSCanto™ II,§ BD™ LSR II, and BD FACSCalibur™ flow cytometers
provides fully automated and rapid sample acquisition from either 96- or 384-well microtiter plates.
• Under 15-minute acquisition for a 96-well tray
• Carryover <1% in high throughput mode, <0.5% carryover in standard mode
• User-adjustable mixing, wash, and acquisition parameters
• Adaptable to a wide range of applications
• Robotic-accessible design
BD Flow Cytometry Instrumentation
BD FACSArray bioanalyzer* Inquire
•Fastmicrotiterplatesampler
•Six-parameterdetection(twoscatterandfourfluorescences)
•Intuitivesoftware
•Digitalsignalprocessingwithupto15,000eventspersecond
•Compact,affordablebenchtopunit
BD FACSCalibur flow cytometer* Inquire
•Theindustrystandardindual-laser,six-parameter,four-colorflowcytometricanalysis
•IncludesBDCellQuest™ProSoftware
•BDFACSCaliburandBDCellQuestProareforInVitroDiagnosticUsewhenusedwithIVDclearedassays
•BDHTSoption
BD FACSCanto II flow cytometer*† Inquire
•Three-laserinstrumentwithtrueeight-colorcapability‡
•Powerfuldigitalsoftwaretomakemulticolorexperimentationfastandsimple
•Resolvesthedimmesteventsbyincreasedsensitivity
•BDHTSoption‡§
BD LSR II flow cytometer* Inquire
•Incorporatesdigitalelectronicsforusewithuptofourfixed-alignmentlasers(488nm,638nm,405nm,andUV)
•Detects18colorsthrougharevolutionarynewopticaldesign
•Flexibleandmodularforfutureupgrades
•BDHTSoption
BD FACSAria cell sorting system* Inquire
•Firstbenchtophigh-speedsorterwithfixed-alignmentcuvetteflowcell
•Cuvetteflowcellforsuperiorfluorescencesensitivity
•Uptothreeair-cooledlasersat488,633,and405nmwavelengths
•Digitalacquisitionratesofupto70,000eventspersecond
•Multicoloranalysisofupto15parameters
•Two-andfour-waybulksortingdevicesforavarietyoftubesizes
•OptionalBD™AutomatedCellDepositionUnit(ACDU)forsortingtoBD™Multiwellplatesormicroscopeslides
instrument features cat.no.
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Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
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Prices are subject to change without notice. All applications are either tested in-house or reported in the literature. See Technical Data Sheets for details.
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