Flow Cytometry Innovative Solutions
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EMD Millipore is a division o Merck KGaA, Darmstadt, Germany
Innovative Solutions orFlow Cytometry Analysis
Optimized Kits and Reagents
http://www.millipore.com/?cid=BIOS-S-EPDF-1227-1108-DS -
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Flow cytometry is an essential tool or in-depth cell
analysis. With the capacity to simultaneously
measure multiple parameters on hundreds o
individual cells per second, ow cytometry oers
greater speed, precision, and detail than most other
cell analysis methods available to scientists today.
Integrated products rom EMD Millipore will help
streamline your workow. Our complete benchtop
ow cytometry solution includes our instruments,
assays, and sotwareas well as the cell
isolation and culture tools to prepare your samples.
With a ow cytometry solution right in your lab,
youll experience superior perormance, higher
quality data and aster progress rom hypothesis to
results.
Guava integratedow cytometry solutions
Whats inside...
Cell Health Cell Counting & Viability
Cell Cycle
DNA Damage
Mitochondrial Analysis
Apoptosis
Cell Signaling MAPK Pathway
EGFR Pathway
PI3/Akt/mTOR Pathway
Jak/STAT Pathway
Chemokine
Stem Cells Embryonic Stem Cell
(Human/Mouse)
Neural Stem Cell
(Rodent)
Immunology Regulation T-Cell
Phenotype Markers
Milli-MarkConjugated
Antibodies
Componentso the guava
Flow CytometrySolution Instruments: easyCyte
Flow Cytometers
Sotware
Kits & Reagents
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Kits and Reagents
FlowCellect Kits and
Milli-Mark Conjugated
Antibodies FlowCellect Kits
Milli-Mark Conjugated
Antibodies
Many researchers invest time
optimizing and validating antibodies
or ow cytometry, only to discover
that these antibodies do not perorm
well when multiplexed together,
because o intererence rom the
matrix or rom other antibodies.
EMD Millipores FlowCellect kits
and Milli-Mark conjugated primary
antibodies are ully optimized
or ast, easy, and accurate
multiparametric ow cytometry.
Weve taken the guesswork out o
assay development so you can ocus
on your results. We optimize and
validate every antibody, making sure
they work together well. All you need
are cells and a research question; our
assay kits will do the rest, and youll
have data beore your cells are ready
to split again.
FlowCellect kits are EMD Millipores
proprietary multiparameter ow
cytometry kits or the analysis
o cellular events and/or cell
phenotypes. Each kit has unique
combination o directly conjugated
antibodies, and/or uorescent dyes
and protein reporters to monitor
changes in protein expression and
posttranslational modifcation. The
kits also contain complete buer
sets, protocols and pre-defned gate
settings. They are ully optimized or
plug-and-play cellular analysis on
guava instruments and other ow
cytometers.
Using a our-step validation process
to develop our FlowCellect kits, weve
eliminated the need to design your
experiment or optimize antibodies
and buers (Figure 2 below).
Milli-Mark antibodies are directly
conjugated primary antibodies
that are validated or ow
cytometry in addition to traditional
applications like Western blotting
and immunocytochemistry (see
Figure 1B or an example o antibody
validation). Because o their
extensive crossplatorm validation,
Milli-Mark antibodies are valuable,
convenient building blocks with
which you can confgure your own
assays.
The antibodies used in
each kit are careully
selected by reviewing
many publications.
Antibodies are conjugated
to compatible
uorophores that will
ensure less uorescence
spectrum overlap.
Antibodies are screened
primarily by Western blot
to determine specifcity,
then validated or ow
cytometry using a
secondary antibody.
The antibody conjugates
are optimized to provide
the best signal-to-noise
ratio when multiplexing.
Figure 2. FlowCellect Kit Four-step Validation Process.
Antibody Selection
and Assay Design
Antibody Conjgation
and Testing
Antibody Component
Validation
Mltiplexing,Stability, Perormance
Claims
Figure1. (A) Mouse embryonicstem cells and fbroblastslabeled with Oct-4 (green),SSEA-1 (red), and Hoechstnuclear stain (blue). (B) Humanembryonic stem cell lysate.
Advantages
Multiplexingcapabilities
Easytouse,withfewerincubationandwashsteps
Fullyvalidatedandconcentration-optimized,guaranteed
to work in ow cytometry
B.
A.
OCT-4antibody
SSEA-4antibody
250
130
100
70
55
35
27
15
10
250
130
100
70
55
35
27
15
10
44kDa
70kDa
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Cell Health
Knowing the perormance profle o your cells prior to
running your bioassay can mean the dierence between
valid assay results and wasted reagents, lost time and
discarded data.
Monitoring key indicators o cell health and perormance,
such as the apoptotic raction and stage o apoptosis,
viability, cell cycle, cell counts, transection efciency, and
target expression levels, helps establish uniorm standards
o cellular perormance across long-term research studies.
These standards can be applied to a wide range obioassays. Whether you are establishing screen/no screen
criteria or high throughput screening, monitoring and
optimizing bioreactor conditions, or eliminating sources o
assay variability, consistent monitoring o your cell model
improves your bioassay perormance and productivity.
Cell Conting and ViabilityCell counting and viability assessments can be used in avariety o applications, such as cytotoxicity studies, PBMC
counting and rapid apoptosis assessment.
gava ViaCont Assay Kits
The guava ViaCount assay is ast becoming the new
standard or viability and cell counting. In this simple
no-wash, mix-and-read-assay, you can accurately obtain
absolute total cell counts, perorm viability assessments
and determine apoptotic percentagesall rom tiny
samples. Youll enjoy several advantages over traditional
methods, including greater accuracy, reproducibility and
speed.
Advantages
Assays:
Simpleno-wash,mix-and-readprocedure
Countsupto10timesfasterthanmanualmethods
Morereproduciblethantraditionaltests
Samples:
Usessmallsamplesintubesora96-wellplate
Handleslow-densityandsmall-volumecellsamples
Workswithadherentorsuspendedcellsandmammalianand insect cells
Description Qty Catalog No.
Guava ViaCount Reagent Kit
The new standard or cell counting and viability assessment.
100tests 4000-0040
600tests 4000-0041
Guava ViaCount Flex Reagent Kit
Non-mammaliancelllines(ie.SF9insectcells).
Lowdensitycellsamples(~104 cells/mL).
Cell lines that stain heterogeneously.
100tests 4500-0110
500tests 4700-0060
Guava ViaCount Cell Dispersal Reagent Kit (CDR)
Uses enzymes to gently disaggregate clumped cells in suspension, improve the accuracy
and precision o cell counts.
100tests 4700-0050
The blue population o cells show a signifcant amount o
annexin V staining (right plot), indicating that intermediate
levels o staining with the viability dye correlates with apoptosis.
Discover the power o ow cytometry
or multiparametric cell health analysis
Viability Stain
Nucleated
Cells
Live
Apoptotic
Dead
Annexin V
Live
Apoptotic
Dead
104
103
102
101
100100 101 102 103 104
104
103
102
101
100100 101 102 103 104
ViabilityS
tain
gava ViaCont Assay Kits
http://www.millipore.com/catalogue/item/4000-0040?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4000-0041?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0110?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4700-0060?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4700-0050?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4700-0050?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4700-0060?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0110?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4000-0041?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4000-0040?cid=BIOS-S-EPDF-1227-1108-DS -
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Cell CycleThe cell cycle can be divided into two distinct stages. Thefrst stage is interphase which consists o the G1, S, and
G2 phases, in which cells are active, growing, and DNA is
being replicated. The second is M phase, also known as the
mitotic phase, in which cell division takes place.
Cell cycle phase distributions can be used to assess cell
health, prolieration, as well as the potential mechanism
o antineoplastic agents. For example, measuring the
population o S phase cells can reect the amount o newly
synthesized DNA. Also, distinguishing cells in G2 rom M
phase cells can help identiy cells undergoing mitosis.
Flow cytometry analysis o cell DNA content has been
one o the best and most popular tools or researchers
to be widely used or the estimation o cell cycle phase
distribution. However, the limitation o single-marker
analysis, such as a DNA dye only, is that cells within
each phase cannot accurately be determined without
mathematical interpolation using analysis sotware.
FlowCellect Cell Cycle KitsEMD Millipore has developed and optimized two bivariate
cell cycle analysis kits using phase-specifc antibodies
in addition to a DNA dye. Bivariate analysis will not only
reveal the cell distribution within a particular phase o
cell cycle, but can also enable the researcher to elucidate
mechanisms o cell cycle regulation, without sophisticated
sotware modules or algorithms.
Advantages
Quantitative
measurements opercentage o cells withineach cell subpopulation
Minimalassaydevelopment needed
Includesalloptimizedow cytometry antibodiesand buers
Nospeciccellcycleanalysis sotware required
Discover the power o ow cytometry
or multiparametric cell cycle research.
FlowCellect Bivariate Cell Cycle Kit
or DNA Replication Analysis
Investigate DNA replication in the S phase with high
accuracy and confdence. The kit includes a directly
conjugated Anti-BrdU Alexa Fluor 488 antibody plus
a DNA dye (propidium iodide). BrdU incorporation is a
widely accepted method o measuring DNA replication
and kinetics o cell cycle progression. The percentage o
BrdU labeled cells is a reliable estimate o the S phase
compartment, and labeled cells can then be ollowed
through the cell cycle.
Detection o DNA replication
by analysis o S phase cells.
Bivariate ow cytometric
analysis using BrdU Alexa
Fluor 488 conjugate can
distinguish S phase cellswith great accuracy, not only
based on their dierence
in DNA content rom G1 or
G2/M cells but also as having
incorporated BrdU.
G=24%
(-BrdU, 1X DNA content)
S=72%
(BrdU, 1-2X DNA content)
G2/M=4%
(-Brdu, 2X DNA content)
FEATuRED PRODuCT
Cell Growth
Cell Prepares
for Division
DNA Synthesis
Mitosis
Cells that
cease dividing
INTERPH AS
E
Cy
tokinesis
Propidium Iodide(x1000)
BrdU
104
103
102
101
1000 1 3 5 7 9
S Phase
G1 G2/M
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FlowCellect Bivariate Cell Cycle Kit
or G2/M Analysis
Investigate the G2/M phase transition with high accuracyand confdence. The phosphorylation o histone H3 at
Ser10correlateswiththeG2toMphasetransitionandis
a prerequisite or chromatin condensation at mitosis. At
the end o mitosis, histone H3 is rapidly dephosphorylated
and remains unphosphorylated throughout the remainder
ofinterphase.Therefore,phospho-histoneH3(Ser10)isa
reliable, specifc marker o M-phase cells.
Nocodazole treatment increases percentage o cells in M phase.
Cell were either treated with 100 m Nocodazole (test sample)
or let untreated (control) overnight at 37 C. By plotting the
phosohorylation o histone H3 at Ser10 (y axis) versus DNA
content (x axis), an increase in the proportion o G2/M cells
was observed indicating that mitotic cells have accumulated
ater treatment. Apporomately 2% o cells reside in M phaseunder normal conditions in Jurkat cells, but when treated cell
population increased 18%.
PE (x1000)
Propidium Iodide
Control (untreated) Nocodazole treated
pH3-A
lexa
Fluor488
p
H3
104
103
102
101
1000 1 3 5 7 9
PE (x1000)
pH3-A
lexa
Fluor488
104
103
102
101
1000 1 3 5 7 9
3% cellsin M phase
18% cellsin M phase
gava Cell Cycle Assay
This kit uses the nuclear DNA stain, propidum iodide (PI), to
measurecellcycle.Restingcells(G0/G1)containtwocopies
o each chromosome. Cycling cel ls synthesize chromosomal
DNA (S phase), which results in increased uorescence
intensity. When all chromosomal DNA has doubled (G2/M
phase), cells uoresce with twice the intensity o the initial
population.
Description Qty Catalog No.
Bivariate Cell Cycle Kit or DNA Replication Analysis Anti-BrdU / Propidium Iodide Solution 25 tests FCCH025102
BivariateCellCycleKitforG2/MAnalysisAnti-phospho-HistoneH3(Ser10)/PropidiumIodideSolution 25 tests FCCH025103
guava Cell Cycle Reagent Propidium Iodide Solution 100tests 4500-0220
Cell Cycle Phases:
G1 = 57%
S = 19%G2 = 15%
M = 3%
FlowCellect Cell Cycle Kits
P1 (x1000)
pH3-Alexa
Fluor488
104
103
102
101
1000 1 3 5 7 9
M
G2
SG1
FlowCellect Bivariate Cell Cycle Kit or G2/M
Analysis (Application)
FEATuRED PRODuCT
Discrimination between G2 and M phase cells by measuring
the phosphorylation o histone H3 on Ser10. Histone H3 is
constitutively phosphorylated at Ser10 during metaphase.
http://www.millipore.com/catalogue/item/FCCH025102?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH025103?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0220?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0220?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH025103?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH025102?cid=BIOS-S-EPDF-1227-1108-DS -
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FEATuRED PRODuCTDNA DamageSignaling PathwayInvestigating the DNA damage signaling pathway is an
important area or genome health and cancer research.
Evidence suggests there is a direct correlation between
DNA damage and cell cycle. Cells that are deective in
DNA damage pathways can cause cancer because they
lack the ability to sense and repair the damage, leading to
genetic instability and ultimately uncontrolled cell growth.
The main kinase activated in response to double-stranded
DNA breaks is ATM or Ataxia telangiectasia mutated
kinase. ATM is a member o the phospho inositide 3-kinase
(PI3K)-related Ser/Thr protein kinase amily. Inactive ATM
exists as a dimer but quickly dissociates and becomes
phosphorylatedonSerine1981inresponsetoionizing
radiation. Once activated, ATM phosphorylates a number
o downstream actors, including P53, CHK2, SMC1, NBS1,and Histone H2A.X.
Description Qty Catalog No.
Mlticolor DNA Damage Response KitAnti-p-SMC1(S957)-AlexaFluor488/Anti-pATM(S1981)-PE/Anti-pHistoneH2A.X(S139)-PerCP
25 Tests FCCH025104
DNA Damage Histone H2A.X Dal Detection KitAnti-pHistoneH2A.X(Ser139)-PerCP/Anti-HistoneH2A.X-FITC
25 tests FCCS025153
Cell Cycle Checkpoint H2A.X DNA Damage KitAnti-pHistoneH2A.X(Ser139),cloneJBW301-AlexaFluor488/PropidiumIodideSolution
25 tests FCCH025142
Cell Cycle Checkpoint ATM DNA Damage KitAnti-pATM(Ser1981),clone10H11.E12-AlexaFluor488/PropidiumIodiden
25 tests FCCH025143
FlowCellect DNA Damage Histone H2A.X
Dal Detection Kit
FlowCellect Histone H2A.X DNA Damage Dual Detection kit
includes two directly conjugated antibodies, a phospho-
specicAnti-phospho-HistoneH2A.X(Ser139)-PerCP
and an Anti-Histone H2A.X-FITC conjugated antibody tomeasure total levels o Histone H2A.X. This two color kit is
designed to detect the extent o Histone H2A.X pathway
activation by measuring H2A.X phosphorylation relative to
the total H2A.X expression in any given cell population. By
doing such, the levels o both the total and phosphorylated
protein can be measured simultaneously in the same cell,
resulting in a normalized and accurate measurement o
H2A.X activation ater stimulation. Moreover, simultaneous
measurement o both total and phospho-Histone H2A.X
confrms target specifcity o the phosphorylation event.
Together, a total and phospho antibody duo perormedin multiplex provides an enhanced and more reliable
detection o the phospho: total ratio within a mixed cell
population. Using this antibody pair provides a sensitive
and valuable tool to study the actors that induce DNA
damage and/or aect DNA repair, and allow one to explore
the linkage between DNA damage, cell cycle checkpoints,
and initiation o apoptosis.
FlowCellect DNA Damage Kits
p53
P
P
MRN
Complex
ATM
ATM
ATM
Ionizing Radiation Changes in
Chromatin Structure
BRCA1
CHK2
P
H2AX
P
P
P
P
P
P
P
P
53BP1 MDC1
SMC1
P
P
Apoptosis
DNA RepairCell-cycle
Checkpoint
Arrest
P
P
A. Unstimulated B. Stimulated
H2AX-FTC (GRN-HLog)
Total H2AX - FITC
Phospho
H2AX
-
PerCP
pH2AX-PerCP
(RED-H_
cg)
H2AX-FTC (GRN-HLog)
pH2AX-PerCP
(RED-H_
cg)
Data below: Unstimulated
HeLa cells are stained with
both phospho-Histone
H2A.X-PerCP and Anti-Histone
H2A.X-FITC (A), where there is
no indication o Histone H2A.Xactivation via phosphorylation,
but only on total H2A.X as
noted by 97.2% o cells.
However, once HeLa cells
were stimulated with 100
M etoposide, simultaneous
measurement o both total
and phospho Histone H2A.X
confrms target specifcity o
the phosphorylation event
as indicated by the double
positive cell population (B)
as indicated by the 2.09%
to 97.15% increase o
double positive staining. Thelevels o both the total and
phosphorylated protein can
be measured simultaneously
in the same cell, resulting in
a normalized and accurate
measurement o H2A.X
activation ater stimulation.
http://www.millipore.com/catalogue/item/FCCH025104?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025153?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH025142?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH025143?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH025143?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH025142?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025153?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH025104?cid=BIOS-S-EPDF-1227-1108-DS -
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Mitochondrial AnalysisMitochondria are critical cellular organelles that produce
90%ofcellularenergy,controlcellsurvivalbyregulatingapoptosis, and produce reactive oxygen species (ROS).
Mitochondrial superoxide generation results in oxidative
stress, damage and cell death by apoptosis or to cellular
energetic decline. Thereore, mitochondrial dysunction
caused by disease or compound treatment has dire
consequences that can result in cell death.
Monitoring impact on mitochondria and related cell health
markers is an important part o drug screening programs,
pathway mapping, apoptosis, and disease research.
Flow cytometry detects multiple markers simultaneously at
various stages o apoptosis, making it a powerul technique
or studying pathways governing cell health and cell death.
FlowCellect Mitochondrial Kits
These kits harness the power o ow cytometry to assess
changes in mitochondrial membrane potential, apoptosis
as measured by Annexin V binding, mitochondrial oxidative
stress, and cell death, using only minimal cellular samples.
The kits may be used with most dual laser ow cytometry
systemsequippedwitha488nmanda644nmlaser.
Advantages
Assay:
Multiplexdetectionwithnooptimizationrequired
Highlyreproducible
Minimalassaydevelopmentneeded
Alloptimizedowcytometryantibodiesandbuffersincluded
Enablesnoviceuserstoperformcomplexanalysis
Samples:
Designedtorun100samplesofhumancells
Discover the power o ow cytometry
or multiparametric mitochondrial
analysis.
Pro-Caspase 8
Pro-Caspase 9
Cyt c
Cyt cAPAF-1
Smac/Diablo
AIF
Endo G
Caspase 8
Caspase 3
Apoptosis
Mitochondria
Apoptosom
e
Nucleus
m MitochondrialPotential Change
Activated Caspase Cascade
ER Stress,DNA Damage,Oxidants
DNA Fragmentation
Chromatin Condensation
BaxBak
t-Bid
IAP
IAP
Bid
MitochondrialSignaling and
Apoptosis
Adapted rom Bayir
and Kagan
Critical Care 2008
12:206.
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FlowCellect MitoDamage Kit
These kits harness the power o ow cytometry to assess
changes in mitochondrial membrane potential, apoptosis
as measured by Annexin V binding, mitochondrial oxidative
stress, and cell death, using only minimal cellular samples.
The kits may be used with most dual laser ow cytometrysystemsequippedwitha488nmanda644nmlaser.
Simltaneosly measres 3 important cell health
parameters:
Changeinmitochondrialpotential(consideredanearly
hallmark o apoptosis or cell stress)
Phospatidylserinetranslocationtothesurfaceofearly
apoptotic cells (measured by Annexin V binding)
Plasmamembranepermeabilizationorcelldeath
(measured by 7-AAD)
The FlowCellect MitoDamage kit can ths distingish
mltiple poplations:
1. Healthy cells with intact mitochondrial membrane
2. Stressed cells with dissipated membrane potential
without Annexin V or 7-AAD staining
3. Early apoptotic cells with dissipated membrane
potential and Annexin V binding
4. Late apoptotic cells or dead cells with dissipated
membrane potentials
Uninduced 50 M CCCP
MitoSenseRed
Annexin V, CF488A
Red2Fluoresecence(RD2-HLog)
Green Fluorescence (GRN-HLog)
94.4%
0.75% 3.7%
100 101 102 103 10410
0
101
102
103
104
1.1%
Red2Fluoresecence(RD2-HLog)
Green Fluorescence (GRN-HLog)
54.7%
14.6% 27.0%
100 101 102 103 10410
0
101
102
103
104
3.7%
Red2Fluoresecence(RD2-HLog)
Green Fluorescence (GRN-HLog)
0.20%
93.2% 6.6%
100 101 102 103 10410
0
101
102
103
104
0.04%
MitoSenseRed
Red2Fluorescence(RD2-HLog)
Red Fluorescence (RED-HLog)
95.2%
3.2% 1.3%
100
101
102
103
104
100
101
102
103
104
0.3%
Red2Fluorescence(RD2-HLog)
Red Fluorescence (RED-HLog)
0.26%
98.4% 1.3%
100
101
102
103
104
100
101
102
103
104
0.02%
100
101
102
103
104
101
102
103
104
Depolarized Cells
Dead Cells
Live Cells
58.1% 0.08%
41.0% 0.8%
Red2Fluorescence(RD2-HLog)
Red Fluoresecence (RED-HLog)
100
2 M Staurosporine
7-AAD
Annexin V, CF488A
Red2Fluorescence(RD2-HLog)
Green Fluorescence (GRN-HLog)
0.16%
95.2% 3.2%100 101 102 103 104
100
101
102
103
104
1.4%
Red2Fluorescence(RD2-HLog)
Green Fluorescence (GRN-HLog)
0.06%
70.5% 28.2%100 101 102 103 104
100
101
102
103
104
1.2%
Red2Fluorescence(RD2-HLog)
Green Fluorescence (GRN-HLog)
0.10%
93.9% 4.8%100 101 102 103 104
100
101
102
103
104
1.2%
7-AAD
Dot plots depicting Jurkat cells
stained using the MitoDamage
kit. Jurkat cells uninduced,
induced to apoptosis with
2 M staurosporine or with
50 M CCCP, then stained
using the MitoDamage kit.
Plots show the percentage o
positive cells or:
1st row: Apoptosis (Annexin
V binding) and mitochondrial
membrane potential change
2nd row: Cell death and
mitochondrial membrane
potential change
3rd row: Apoptosis and cell
death.
Data reports that 2 M
staurosporine induces
apoptosis in Jurkat cells, and
that 50 M CCCP depolarizesthe mitochondrial membrane,
but neither condition is
sufcient or cell membrane
permeabilization and death.
Kit Component Laser Ber Pack
MitoSense Red Dye Red10XAssay
Buer HSCAnnexin V-CF488A Blue
7-AAD Blue
FEATuRED PRODuCT
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FlowCellect MitoPotential Red Kit
Simultaneous analysis o mitochondrial membrane
potential along with cell death provides key inormation
or drug discovery, cancer and toxicology studies as well
as disease-induced apoptosis. This kit uses MitoSense
Red (a red laser-excitable dye) to monitor mitochondrial
membrane potential changes in early apoptosis, and
7-AAD (a live-cell-impermeant DNA intercalator) to
simultaneously monitor cell membrane permeability
changes in late apoptosis and necrotic cell death.
FlowCellect MitoLive Kit
Incldes:
MitoSense Red, a uorescent cationic dye that
accumulates in the mitochondria and is responsive to
mitochondrial potential changes
Calcein acetoxymethylester (calcein-AM) a non-
uorescent, cell-permeant compound that is hydrolyzed
by intracellular esterases into the uorescent anion
calcein and provides a measure o cellular vitality.
The simultaneous use o the reagents enables researchers
to obtain inormation on early and late apoptosis in one
simple assay.
FlowCellect MitoStress Kit
Incldes:
MitoSOX Red, a live-cell-permeant, uorogenic
dye which targets the mitochondria and reacts with
superoxide radicals and uoresces yellow/red
AnnexinVconjugatedtoCF647whichbindsto
phosphatidylserine (PS) on the surace o apoptotic cells
The simultaneous use o these reagents enables researchers
to obtain inormation on oxidative stress and apoptosis in
one simple assay.
FlowCellect Cytochrome c Kit
Includes a directly labeled anti-Cytocrome cFITC antibody
and Anti-IgG1-FITC isotype control. Viable or live cells will
demonstrate higher levels o Cytochrome c staining while
apoptotic cells which have released their Cytochrome c
rom the mitochondria to the cytoplasm will demonstrate
reduced staining intensity. The FlowCellect Cytochrome c
ow cytometry kit is a simple, gentle, and ast method to
assess levels o Cytochrome c in mitochondria, providing a
valuable tool or assessing pro-apoptotic signaling and the
efcacy o pro-apoptotic anti-cancer agents in cells.
Description Qty Catalog No.
FlowCellect MitoPotential Red Kit Two dyes or measuring cell death and mitochondrial membrane potential
MitoSense Red (Red Laser) / 7-AAD (Blue Laser)
100tests FCCH100105
FlowCellect MitoDamage Kit
Three dyes to assess mitochondrial potential, stress, and cell death
Mitosense Red (Red) / Annexin V-CF488A (Blue) / 7-AAD (Blue Laser)
100tests FCCH100106
FlowCellect MitoLive Kit
Two dyes to measure mitochondrial health and cell vitality
Mitosense Red (Red) / Calcein-AM (Blue Laser)
100tests FCCH100107
FlowCellect MitoStress Kit
Understanding the regulation o apoptosis and oxidative stress.
MitoSoxRed(Red)/AnnexinV-CF647(BlueLaser)
100tests FCCH100109
FlowCellect Cytochrome c Kit
Easy way to detect loss o mitochondrial Cytochrome c in cells
Anti-Cytochrome c-FITC (Blue) / Anti-IgG1-FITC (Blue Laser)
100tests FCCH100110
FlowCellect Oxidative Stress Characterization Kit
Detection o oxidative stress by ow cytometry
Anti-DNP-FITC (Blue Laser)
25 tests FCCH025111
guava Mitochondrial Depolarization Assay Kit
Monitoring changes in mitochondrial membrane potential
JC-1 (Blue Laser) / 7-AAD (Blue Laser)
100tests 4500-0250
FlowCellect Mitochondrial Kits
http://www.millipore.com/catalogue/item/FCCH100105?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100106?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100107?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100109?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100110?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH025111?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0250?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0250?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH025111?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100110?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100109?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100107?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100106?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100105?cid=BIOS-S-EPDF-1227-1108-DS -
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Early Mid Late
Guava Nexin Multicaspase Guava TUNEL
Annexin Red Caspase 3/7, 8, 9 Assay
MitoPotential Red Dual Caspase
MitoDamage
MitoStress
MitoLiveCytochrome c
c
cc
c
c+
PS
PS
PS
PS
PS
PS
c+c+
c+
c+
c+
G...C
G...C
C...G
A...T
G...C
C...G
A...T
A
G
G...C
G...C
C...G
A...T
G...C
C...G
A...T
A
G
G...C
G...C
C...G
A...T
G...C
C...G
A...T
A
G
TdT
+
BrdUTP
TRITC-
Anti-BrdU
DNA strand
breaks de to
apoptosis
Add BrduTP
to 3OH ends
Antibody labeled
break sites
ApoptosisCells respond to specifc apoptotic signals by initiating
intracellular processes that result in characteristic
physiological changes. Among these changes are
externalization o phosphatidylserine to the cell surace,
depolarization o mitochondrial membranes, cleavage
and degradation o specifc cellular proteins, compaction
and ragmentation o nuclear chromatin, loss o cell
membrane integrity, and cellular shrinkage. Suppression or
enhancement o apoptosis is known to cause or contribute
to diseases such as cancer and diabetes, making the
apoptotic pathway a popular drug target.
Because apoptosis is a dynamic event, and the time period
during which cells exhibit apoptosis markers is variable andshort, ow cytometry is an ideal technique or tracking
cells through apoptosis. Our easy-to-use kits enable you to
examine cells at each o the various stages o apoptosis.
Early Apoptosis Flow Cytometry Kits
Two separate dyes identiy a broad spectrum o
apoptotic and non-apoptotic cells: Annexin V binds to
phosphatidylserine on the external membrane o apoptotic
cells, while 7-AAD permeates and stains DNA o late-stage
apoptotic and dead cells.
Advantages
Twodyestrategy:Detectvariousstagesofapoptosiswithin a one assay
Mix-and-readassay:Getstandardizedresultsevenwithmultiple users
All-in-one-kit:Spendlesstimebeforeanalysis
CompatiblewithpairingwithFITCorPEprobesorotherprobes in the green or yellow channels
Probewithothermarkersingreenandyellowchannels
with the FlowCellect Mitochondrial kits
Mid-Apoptosis Flow Cytometry Kits
Caspase activity is measured using a FLICA (uorescent
labeled inhibitor o caspase) reagent, supplemented by
a nuclear DNA stain 7-AAD, which evaluates membrane
integrity and cell viability. The assays are available in two
orms, sulorhodamine (SR) and carboxyuorescein (FAM),
giving greater exibility in assay design as well as the
capacity to multiplex caspase assays.
Late Apoptosis Flow Cytometry Kits
The guava TUNEL assay detects apoptosis-induced DNA
ragmentation through a quantitative uorescence assay.
Terminal deoxynucleotidyl transerase (TdT) catalyzes the
incorporation o bromo-deoxyuridine (BrdU) residues into
the ragmented nuclear DNA at the 3-hydroxyl ends. A
TRITC-conjugated anti-BrdU antibody then labels theseDNA ragments. The assay distinguishes two populations:
non-apoptotic cells (TUNEL-negative) and apoptotic cells
(TUNEL-positive).
Discover the power o ow cytometryor multiparametric apoptosis analysis.
Live/Healthy
(non-committed)cells
FLICA(-)7-AAD(-) FLICA(+)7-AAD(-) FLICA(+)7-AAD(+) FLICA(-)7-AAD(+)
Early/Mid-stage
(committed)apoptotic
cells
Late stage
apoptotic/dyingcells
Dead cells
C+C+C+
C+
C+C+ C+
C+PI+PI+
PI+
PI+PI+
PI+
-
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FlowCellect Annexin Red Kit
In the early stages o apoptosis, phosphatidylserine
molecules, which can bind to Annexin V, move rom the
inner leaet, to the outer leaet o the plasma membrane.
A rapid, sensitive, and convenient assay to monitor early
and late apoptosis, the FlowCellect annexin red kit includesrecombinant Annexin V conjugated to the red sensitive
dyeCF647,and7-AAD(alivecell-impermeantdye)to
measure cell membrane integrity. Ater staining cells with
FlowCellect Annexin Red kit, three populations o cells can
be identifed in this assay:
Non-apoptotic cells: Annexin V(-) and 7-AAD(-)
Early apoptotic cells: Annexin V(+) and 7-AAD(-)
Late-apoptotic or dead cells: Annexin V (+) and 7-AAD(+)
Two Dyes to distinguish early Apoptosis rom later stages
Dot plots depicting Jurkat cells stained using the FlowCellect Annexin Red kit. Jurkat cells were untreated (Plot A), treated with
0.1 M (Plot B) or with 3 M staurosporine (Plot C), and then stained using the FlowCelllect Annexin Red kit. The percentage o
apoptotic cells increased rom 36% to 92% in response to a 30-old increase in staurosporine concentration; however, only a small
raction (< 10%) o the cells showed evidence o cell death.
Kit Component Laser Ber Pack
AnnexinV-CF647 Red 10XAssay
Buer HSC7-AAD Blue
FEATuRED PRODuCT
7-AAD
B. C.
Annexin V
A.
Red2Fluorescence(RD2-HLog)
2% 2%
85% 11%
Red2 Fluorescence (RED2-HLog)
100 101 102 103 104100
101
102
103
104
100
101
102
103
104
Red2Fluorescence(RD2-HLog)
4%
60% 36%
Red2 Fluorescence (RED2-HLog)
100 101 102 103 104100
101
102
103
104
Red2Fluorescence(RD2-HLog)
7%
5% 92%
Red2 Fluorescence (RED2-HLog)
100 101 102 103 104
3 M Staurosporine0.1 M StaurosporineUninduced
PS = phosphatidylserine
Apoptotic Cell MembranesHealthy Cell Membranes
Annexin CF647
PSPS
PS
PS
PSPS
PS
PS
= 7aad
FlowCellectAnnexinRedKit(AnnexinV-CF647Reagent/7-AAD) 100tests FCCH100108
Guava Nexin Reagent (Annexin V-PE/ 7-AAD) 100tests 4500-0450
Guava TUNEL Reagent Kit (Anti-BrdU -TRITC/ TdT Enzyme/ Br-dUTP) 100tests 4500-0121
FlowCellectBcl-2ActivationDualDetectionKit(Anti-Bcl-2-AlexaFluor488/Anti-pBcl-2(Ser70)-PE) 25 Tests FCCS025108
Guava MultiCaspase SR Kit (MultiCaspase SR reagent / 7-AAD) 100tests 4500-0500
Guava Caspase 3/7 SR Kit (Caspase 3/7 SR reagent / 7-AAD) 100tests 4500-0510
Guava MultiCaspase FAM Kit (Multicaspase FAM reagent / 7-AAD) 100tests 4500-0530
Guava Caspase 3/7 FAM Kit (Caspase 3/7 FAM reagent / 7-AAD) 100tests 4500-0540
Early Apoptosis Kits
Late Apoptosis Kit
Apoptosis Signaling Kit
Mid Apoptosis Kits
For a complete listing visit: www.millipore.com/midapoptosis_kits
Apoptosis Kits
Description Qty Catalog No.
http://www.millipore.com/catalogue/item/FCCH100108?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0450?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0121?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025108?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0500?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0510?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0530?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0540?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025108?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0540?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0530?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0510?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0500?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025108?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0121?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0450?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100108?cid=BIOS-S-EPDF-1227-1108-DS -
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Description Qty Catalog No.
FlowCellect EGFR/MAPK Pathway Activation Detection KitAnti-pEGFR (Tyr1173), AlexaFluor 488
Anti-pERK1/2(Thr202/Tyr204,Thr185/Tyr187)-PE
25 Tests FCCS025101
FlowCellect EGFR RTK Activation Dal Detection KitAnti-pEGFR (Tyr1173), AlexaFluor 488
Anti-EGFR-PerCP
25 Tests FCCS025107
FlowCellect EGFR/STAT3 Pathway Activation Detection Kit
Anti-pEGFR (Tyr1173), Alexa Fluor 488Anti-pSTAT3(Tyr705),AlexaFluor647
25 Tests FCCS025111
FlowCellect Kits: MAPK Pathway
FlowCellect Kits: EGFR Pathway
Cell Signaling
Signal transduction pathways lead to diverse outcomes,such as apoptosis, cell dierentiation, cell growth and cell
prolieration, all o which have been extensively studied
in the process o developing therapies or various cancers
and autoimmune disease. Cross-talk among signaling
pathways adds an extra dimension o complexity when
analyzing physiological consequences o a pathway o
interest. However, there are some key nodes at which
multiple signals are integrated. Multiparametric ow
cytometry provides researchers the power to monitor these
intracellular checkpoints simultaneously, enabling analysis
o complicated cell events.
FlowCellect Cell Signaling KitsThe study o cell signaling has been made easier by
activation status-specifc and phospho-specifc antibodies.
Measuring the activity o cell signaling pathways by ow
cytometry delivers robust, high content inormation in
less time than traditional methods by analyzing multiple
parameters on hundreds o cells per second.
Dal Detection FlowCellect Kits With Pairs oTotal and Phospho-Specic Antibodies
EMD Millipores FlowCellect Dual Detection kits are a
series o ow cytometry products which include a pair
o antibodies that bind to the same protein; one to
detect total protein expression and another to detect
the phosphorylated orm o the same target. Using
two parameter analysis, we can achieve target specifc
detection o phosphorylation and, by doing so, eliminate
alse positives while enhancing the signal to noise ratio.
FlowCellect Cell Signaling Kits With Directly
Conjgated, Phospho-Specic Antibodies
Determine the eect o mechanical and chemical reagents
that can induce DNA damage, discern multiple pathway
activation and cross talk in a time-dependent manner,
or study the correlation between pathway activation and
changes in cell unction and health.
Discover the power o ow cytometry
or multiparametric cell signaling research.
Description Qty Catalog No.
FlowCellect PI3K/MAPK Dal Pathway Activation and Cancer Marker Detection KitAnti-pErk1/2(Thr202/Tyr204,Thr185/Tyr187)-PE
Anti-phospho-Akt1/PKBa (Ser473), Alexa Fluor 488
Anti-KI-67-PerCP
25 Tests FCCS025100
FlowCellect EGFR/MAPK Pathway Activation Detection KitAnti-pEGFR (Tyr1173), AlexaFluor 488
Anti-pERK1/2(Thr202/Tyr204,Thr185/Tyr187)-PE
25 Tests FCCS025101
FlowCellect MAPK Activation Dal Detection KitAnti-pERK1/2(Thr202/Tyr204,Thr185/Tyr187)-PE
Anti-ERK1/2-AlexaFluor647
25 Tests FCCS025106
FlowCellect p38 Stress Pathway Activation Detection KitAnti-pP38(Thr180/Tyr182),AlexaFluor488Anti-pATF2(TThr69/71),AlexaFluor647
25 Tests FCCS025132
AdvantagesAssay:
Ensuresspeciclabelingo targets
Multiplexdetectionwithno optimization required
Enablesnoviceusersto perorm complexanalysis
Samples:
Designedtorun25samples o human cells
http://www.millipore.com/catalogue/item/FCCS025101?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025107?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025111?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025100?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025101?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025106?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025132?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025111?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025100?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025111?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025107?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025101?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025101?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025132?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025106?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025101?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025101?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025100?cid=BIOS-S-EPDF-1227-1108-DS -
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FlowCellect PI3K/MAPK Dal Pathway
Activation and Cancer Marker
Detection Kit
Three antibodies to study cross-talk between the PI3K and
MAPK pathways. The kit uses directly labeled antibodies
against phospho-Akt1/PKBa(Ser473)-AlexaFluor 488 andAnti-phospho-Erk1/2(Thr202/Tyr204,Thr185/Tyr187)-PE
toanalyzesignalingactivationandcrosstalk,plusKi-67
marker-PerCP marker to identiy the prolierative raction.
Together, the antibody trio makes it easy to evaluate the
role these signaling pathways play in prolieration and
dierentiation. The kit also includes Cell Cycle Stop
xationreagenttoimproveKi-67detection.Although
Ki-67ispresentthroughoutmostofthecellcycle,itis
difcult to detect by ow cytometry except during M
phase. Cell cycle stop reagent arrests the cycle at the
Mphase,makingitpossibletoaccuratelydetectKi-67
expression and discern the biological eects o the PI3K/
MAPK cross talk.
FEATuRED PRODuCT
GTP
GDP
Shc
Grb2
Ki 67
PI3-Kinase
Proliferation/
Survival
l
Growth/
Differentiation
Wortmannin
or LY294002
PMA
p85
IGF-1
p110 RAS
Raf-1Akt
P
P
P
P
P
SOS
MEK1/2
Inhibiting
Activating
ERK1/2
P
The cross-talk between the PI3K/Akt and MAPK signaling pathways is evident downstream o IGF in HEK293 cells. Cells are
stimulated at 3 minutes and 5 minutes by Insulin and PMA as shown in A and B, respectively. As previously indicated, although
Insulin initially activates both pathways independently, the activation o the PI3K pathway indicated by the phosphorylation o Akt
and ERK1/2 inhibits the activation o ERK. The dot plots illustrate the cross-talk that exists between these two signaling pathways.This is demonstrated by the sharp decrease in ERK phosphorylation between 3 minutes and 5 minutes (see bar graphs; C, D).
pAKT-Alexa 488
pAKT
Untreated
A. 3 Minute Stimulation C. HEK203 - 3 Minute Stimulation
D. HEK203 - 5 Minute StimulationB. 5 Minute Stimulation
Insulin Treated PMA Treated
pERK-PE
pERK
104
104
103
103
102
102
101
101100
100
pAKT-Alexa 488 pAkt pErk
Untreated100 nM insulin 3 min100 g/mL PMA 3 min
Ki67
010
pERK-PE
%o
fcells
104
104
103
103
102
102
101
101100
100
pAKT-Alexa 488
pERK-PE
104
104
103
103
102
102
101
101100
100
pAKT-Alexa 488
pERK-PE
104
104
103
103
102
102
101
101100
100
pAKT-Alexa 488
pERK-PE
104
104
103
103
102
102
101
101100
100
pAKT-Alexa 488
pERK-PE
104
104
103
103
102
102
101
101100
100
2030405060708090
100
pAkt pErk
Untreated100 nM insulin 3 min100 g/mL PMA 3 min
Ki67
010
%o
fcells
2030405060708090
100
-
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FlowCellect Kit EGFR RTK Activation
Dal Detection Kit
EGF pathway activation through EGFR plays a key role
in the regulation o essential cellular processes, and
abnormality o EGF signaling is linked to cancer. EMD
Millipores FlowCellect EGFR RTK Activation Dual Detectionkit includes two directly conjugated antibodies, a phospho-
specifc Anti-phospho-EGFR (Tyr1173)-Alexa Fluor 488
and an Anti-EGFR-PerCP conjugated antibody to measure
total levels o EGFR. This two color ow cytometry kit is
designed to detect the extent o EGF pathway activation
by measuring the EGFR phosphorylation in relative to the
total EGFR expression in any given cell population. By
doing such, the levels o both the total and phosphorylated
protein can be measured simultaneously in the same cell,
resulting in a normalized and accurate measurement o
EGFR activation ater stimulation. Moreover, simultaneous
measurement o both total and phospho-EGFR confrms
target specifcity o the phosphorylation event. Together,
a total and phospho antibody duo perormed in multiplex
provides an enhanced and more reliable detection o the
phospho:total ratio within a mixed population.
FEATuRED PRODuCT
EGF
EGFR
Ras
ERK
PI3KJAK
AKTSTAT
P P
mTOR
EGFR-PerCP
Total EGFR
A. Isotype Control(No EGF Stimulation)
B. Total and pEGFR(No EGF Stimulation)
C. Isotype Control(EGF Stimulation)
D. Total and pEGFR(EGF Stimulation)
pEGFR-Alexa
Fluor488
pEGFR
104
104
103
103
102
102
101
101100
100
EGFR-PerCP
pEGFR-Alexa
Fluor488
104
104
103
103
102
102
101
101100
100
EGFR-PerCP
pEGFR-Alexa
Fluor488
104
104
103
103
102
102
101
101100
100
EGFR-PerCP
pEGFR-Alexa
Fluor488
104
104
103
103
102
102
101
101100
100
98% 88.6%
5.4%0.5%
5.5%
1.0%
0.6%
0.7%
0% 0.3%
0.5%
0% 1.5%
0.2%
98.2
99%
Dual Parameter Analysis o
Total and Phospho EGFR onA431Cells As illustrated in
dot plots A and B, untreated
A431cells stained with an
isotype control (A) and both
pEGFR-Alexa Fluor 488
and Anti-EGFR-PerCP (B),
respectively, only indicated
EGFR expression on total EGFR
as noted by 89% o cells.
However, once A431cells were
stimulated with 100 ng/
mL EGF, simultaneous
measurement o both total
and phospho EGFR confrms
target specifcity o thephosphorylation event as
indicated by the double
positive cell population (D) as
indicated by the 5% to 98%
increase o double positive
staining. A431 stimulated
cells showed no activity
when stained with an isotype
control (C).
-
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FlowCellect Chemokine Receptor
Srace Expression Qantication Kits
EMD Millipore oers 11 GPCR surace identifcation ow
cytometry kits. Flow cytometry provides high quality,
reproducible data in ar less time than traditional methods
or chemokine receptor research and avoids the hazard and
expense o radioligand binding assays.
Our FlowCellect GPCR identifcation kits can be used to
identiy and quantiy GPCRs on the surace o any cells.
The kits detect GPCR expression using a GPCR-specifc
antibody validated or ow cytometry. Also included arepositive and negative control cells with well-characterized
receptor expression levels or the purposes o quantitative
extrapolation.
Advantages
Assay:
Includespharmacologicallycharacterizedpositiveandnegative control cells
Sameaccuracyasradioactiveassays,withoutthehazards.
Abilitytoidentifyhigh,medium,andlowexpressingcellcultures during the clonal selection process.
HighreproducibleresultsobtainedbyusingEMDMillipores well-characterized ChemiScreen GPCR cell
lines as assay controls
Samples:
Sufcientreagentstorun100samplesofhumancells.
For a complete listing o kits visit: millipore.com/fowcytometry.
Description Qty Catalog No.
FlowCellectChemokineReceptorCCR2BSurfaceExpressionQuanticationKit 100tests FCCR200411
FlowCellectChemokineReceptorCCR4SurfaceExpressionQuanticationKit 100tests FCCR400413
ChemokineReceptorCCR6SurfaceExpressionQuanticationKit 100tests FCCR600414
FlowCellectChemokineReceptorCCR7SurfaceExpressionQuanticationKit 100tests FCCR700415
FlowCellect PI3K/MAPK Dal Pathway Activation and Cancer Marker Detection KitAnti-phospho-Erk1/2(Thr202/Tyr204,Thr185/Tyr187)-PE/Anti-p-Akt1/PKBa (Ser473)- Alexa Fluor 488 /
Anti-KI-67-PerCP
25 Tests FCCS025100
FlowCellect PI3K Activation Dal Detection KitAnti-phospho-Akt(Ser473)AlexaFluor488/Anti-Akt/PKB-AlexaFluor647
25 Tests FCCS025105
FlowCellect PI3K-mTOR Signaling Cascade Mapping KitAnti-p-RibosomalProteinS6(Ser235)-PerCP/Anti-p-Akt1/PKBa (Ser473)-Alexa Fluor 488
25 Tests FCCS025210
FlowCellect EGFR/STAT3 Pathway Activation Detection KitAnti-pEGFR(Tyr1173),AlexaFluor488/Anti-pSTAT3(Tyr705),AlexaFluor647
25 Tests FCCS025111
FlowCellect Mlti-STAT Activation Proling KitAnti-pSTAT1(Tyr701)-PerCP/Anti-pSTAT3(Tyr705)-Alexa488/Anti-pSTAT5A/5B(Tyr694/Tyr699)-PE
25 Tests FCCS025550
FlowCellect STAT1 Activation Dal Detection KitAnti-p-STAT1(Tyr701)AlexaFluor488/Anti-STAT1-PerCP
25 Tests FCCS025142
FlowCellect STAT3 Activation Dal Detection KitAnti-p-STAT3(Tyr705)AlexaFluor647/Anti-STAT3-AlexaFluor488
25 Tests FCCS025143
FlowCellect Bcl-2 Activation Dal Detection KitAnti-Bcl-2 Alexa Fluor 488 Antibody
Anti-pBcl-2(Ser70)PE
25 Tests FCCS025108
Chemokine Receptor Kits
PI3/ Akt/ m-TOR Pathway
Jak/ STAT Pathway
Mltiple Pathway
Apoptosis Signaling Pathway
Description Qty Catalog No.
FlowCellect Cell Signaling Kits
FlowCellect Src Activation Dal Detection KitAnti-pSrc(Tyr416)-AlexaFluor488
Anti-Src-AlexaFluor647
25 Tests FCCS025154
FlowCellect PLC-1 Activation Dal Detection KitAnti-pPLC-1 (Tyr783) Alexa Fluor 488
Anti-PLC-1-PE
25 Tests FCCS025145
http://www.millipore.com/flowcytometry/flx4/flowcytometry?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/flowcytometry/flx4/flowcytometry?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCR200411?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCR400413?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCR600414?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCR700415?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025100?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025105?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025210?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025111?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025550?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025142?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025143?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025108?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025154?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025145?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/flowcytometry/flx4/flowcytometry?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCR700415?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCR600414?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCR400413?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCR200411?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025108?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025145?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025154?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025143?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025142?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025550?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025111?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025210?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025105?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025100?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025111?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025100?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCS025108?cid=BIOS-S-EPDF-1227-1108-DS -
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FlowCellect Hman ESC (Oct4) Nclear Marker Characterization Kit
hOCT4-Alexa 488 / SSEA4-PE / SSEA1-PE/CY5
25 Test FCHEC25102
FlowCellect Hman ESC (HESCA-1) Srace Marker Characterization KitHESCA1-FITC / SSEA4-PE / SSEA1-PE/CY5
25 Tests FCHEC25104
FlowCellect Hman ESC (TRA-1-60) Srace Marker Characterization KitTRA-1-60-FITC/SSEA4-PE/SSEA1-PE/CY5
25 Tests FCHEC25106
Hman
FlowCellect Rodent NSC Characterization Kit (Neronal Dierentiation)Sox-2 FITC / Nestin-PE / Beta-III-Tubulin-PE/CY5
25 Tests FCRNC25112
Rodent
FlowCellect Mose ESC (Oct4) Nclear Marker Characterization KitmOCT4-Alexa 488 / SSEA4-PE / SSEA1-PE/CY5
25 Tests FCMEC25110
Mose
Stem Cells
Because ow cytometry has the power to characterize
subpopulations o cells within heterogenous cell mixtures;
it is widely used or studying both embryonic stem cells
(ESCs) and induced pluripotent stem (iPS) cells. Flow
cytometry enables researchers to evaluate percentages
o cells expressing specifc markers, to determine culture
quality, and to track gene expression changes during a
dierentiation protocol.
FlowCellect Stem CellCharacterization KitsEMD Millipores FlowCellect stem cell characterization
kits are designed to provide rapid, sensitive assessments
o embryonic and neural stem cell phenotypes at various
stages. The kits use three parameters or accurate
identifcation, enabling the research to triangulate
cellular phenotypes with two complementary positive
markers and one negative marker. The negative antibody
also serves as a stage- and species-specifc or lineage-
specifc marker or dierentiated cells.
Advantages
Assay:
Alloptimized,uorescently-labeledowcytometryantibodies and buers included
Highlyreproducibleresults
Validatedforowcytometry,immunocytochemistry,andWestern blotting
Discover the power o ow cytometry
or multiparametric stem cell analysis.
Embryonic Stem Cell Markers
Stem cell(hESC, mESC)
Oct-4+
SSEA-4+ (human)
TRA1-60+
HESCA+Nanog+
SSEA-1+ (mouse)
Committed cell(ENStem-A)
Oct-4-/+
SSEA-4-
TRA1-60-
HESCA-
Dierentiated
cellsOct-4-
SSEA-4-
TRA1-60-
HESCA-
SSEA-1+/-
Description Qty Catalog No.
Stem Cell FlowCellect Kits
http://www.millipore.com/catalogue/item/FCHEC25102?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCHEC25104?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCHEC25106?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCRNC25112?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMEC25110?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMEC25110?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCRNC25112?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCHEC25106?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCHEC25104?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCHEC25102?cid=BIOS-S-EPDF-1227-1108-DS -
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FlowCellect Hman Embryonic Stem Cell
Characterization Kit
The kit is provides a quick, easy way to track surace marker
expression o hOCT4, SSEA4 and SSEA1. The percentage o
undierentiated human embryonic stem cells in culture is
reected in the percentage cells that express both hOCT4
FlowCellect Hman Embryonic
Stem Cell HESCA-1 Srace Marker
Characterization Kit
This kit provides an easier and quick way to track suracemarker expression o HESCA-1, SSEA4 and SSEA1.
The kit will also help to determine the percentage o
undierentiated human embryonic stem cells in culture
by determining the percentage o cells that express both
and SSEA4 but not SSEA1. This quick test will enable
researchers to determine the multipotency o their cells in
culture as well as see changes in marker expression during
a dierentiation protocol.
HESCA-1 and SSEA4 and not SSEA1. This quick test willenable researchers to test the quality o their cells in
culture as well as see changes in marker expression during
a dierentiation protocol.
FEATuRED PRODuCTS
Representative data o H1 human embr yonic stem cells stained with hOCT4-Alexa 488, SSFA4-PE and SSFA1-PF/CY5
Representative data o H1 human embr yonic stem cells stained with HESCA-1-FITC, SSEA4-PE and SSEA1-PE/CY5.
0%
4.9%
0.1%
95%
10e4
10e3
10e2
10e1
10e0
10e0 10e1 10e2 10e3 10e4
SSEA1-PE/CY5
SSEA4-PE
10e4
10e3
10e2
10e1
10e0
10e0 10e1 10e2 10e3 10e4
HESCA-FITC
SSEA4-PE
10e485.5%
14.4%
0.1%
0%
10e3
10e2
10e1
10e0
10e0 10e1 10e2 10e3 10e4
hOCT4-Alexa488
SSEA1-PE/CY5
B. C.A.
B. C.A.
10e40.1%
4.8%
85.5%
9.6%
10e3
10e2
10e1
10e0
10e0 10e1 10e2 10e3 10e4
hOCT4-Alexa488
SSEA4-PE
10e4
10e3
10e2
10e1
10e0
10e0 10e1 10e2 10e3 10e4
HES
CA-FITC
SSEA1-PE Cy5
10e4
10e3
10e2
10e1
10e0
10e0 10e1 10e2 10e3 10e4
SSEA
1-PECy5
SSEA4-PE
-
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Immunology
The immune system, which mediates the bodys response
to the introduction o oreign material, is made up
o multiple cell types collectively called lymphocytes.
Lymphocyte subtypes include B cells (which secrete
antibodies), cytotoxic T cells, helper T cells (which secrete
cytokines), and natural killer (NK) cells. Characterization o
lymphocyte subtypes and cytokine signaling is essential or
understanding the complex nature o the immune system.
Activation by antigens, suppression o normal immune
activation, and disease states can aect the phenotypes
o lymphocytes. Multiparameteric phenotypic analysisby ow cytometry allows researchers to distinguish one
subpopulation o cells within a heterogeneous mixture,
and thereby enables the study the dynamics o immune
signaling in intact cells.
FlowCellect Immnology KitsEach kit includes multiple optimized uorescent labeled
antibodies and all buers necessary or cell preparation and
analysis. Detailed assay instructions are included to assist in
analysis and to ensure that the correct cell concentration is
obtained during acquisition o sample data.
Advantages
Assay:
Multiplexdetectionwithnooptimizationrequired
Highlyreproducible
Minimalassaydevelopmentneeded
Alloptimizedowcytometryantibodiesandbuffersincluded
Enablesnoviceuserstoperformcomplexanalysis
Samples:
Designedtorun25tests(FlowCellectkits)
Designedtorun100tests(guavakit)
Discover the power o ow cytometry ormultiparametric analysis o theimmune system.
DC
IL-12R
TH1
T-bet
STAT4
STAT1
IL-2R
TRE G
Foxp3
STAT5
IL-21R
TFHTH
Bcl-6
STAT3
IL-4R
TH2
GATA3
STAT6
STAT5a
IL-23R
TH17
RORt
STAT3
IL-12
CD80
CD28
TCR
CD28
CD85
pMHCII
IFN
IL-18
IL-2
IL-4
IL-33
IL-6
IFN
IL-2LT
IL-21
IL-6
TGFIL-23
TGF
IL-2
IL-4
IL-5
IL-13
IL-25
IL-17
IL-17F
IL-22
IL-21
IL-21
IL-17
TGF
IL-10
IL-35
B ZONE
CD62Llo CCR7lo
CCR7lo
High
TCR Binding
Low
TCR Binding
Medium
TCR Binding
CXCR5hi
CD62Lhi CCR7hi T ZONE
EMIGRANTnaive
CD4+ T Cell Dierentiation. Schematic diagram o CD4+ T-cell dierentiation. Five dierent types o CD4+ T-cells can develop rom
a common nave precursor depending on the cytokine environment and interaction with dendritic cells (DC).
-
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FlowCellect Mose Th1/Th17
Intracelllar Cytokine KitEMD Millipores FlowCellect Mouse Th1/Th17 Identifcation
Kit is designed to enable a researcher a quick and easy way
to detect IFN-g and IL-17 expression in mouse Th1 and
Th17 CD4+ T-cells. This kit contains an anti-IFN- antibodyconjugated to PE, an anti-IL17 antibody conjugated FITC
and an anti-CD4 antibody conjugated to PerCP. The kit also
includes optimized a protein transport inhibitor and buers
to aid in identiying Th1 and Th17 CD4+ T-cell subsets in
ex vivo lymphocyte populations or to monitor Th1 and
Th17 CD4+ Tcell dierentiation in culture. In addition, EMD
Millipore has added a fxable viability dye that eliminates
alse positive data resulting rom cytokine staining on dead
cells that can occur in long term cultures. The kit contains
sufcient reagents or 25 3-color tests. Detailed assay
instructions are included to assist in sample preparation
and to ensure that the correct cell concentration is obtained
during acquisition o sample data. This kit is not designed to
be used in the analysis o whole spleen cells in SJL mice.
FEATuRED PRODuCT
Description Qty Catalog No.
FlowCellect Hman FOXP3 Treg Characterization KitAnti-CD3-PE/Cy5/Anti-CD4-FITC/Anti-FOXP3-AlexaFluor647
25 Tests FCIM025118
FlowCellect Hman CD4/CD8 T Cell KitAnti-CD8-FITC, CD4-PE, CD3-PECY5 o cocktail
100Tests FCIM100158
Description Qty Catalog No.
FlowCellect Mose Th1 Intracelllar Cytokine KitAnti-CD4 clone GK1.5-PerCP /Anti-IFN, clone XMG1.2-PE
A quick and easy way to detect IFN-expression in mouse Th1 CD4+ T cell
25 Tests FCIM025123
FlowCellect Mose Th2 Intracelllar Cytokine KitAnti-CD4-PerCP clone GK1.5/ Anti-IL-4, clone 11B11-PE
A quick and easy way to detect IL-4 expression in mouse TH2 CD4+ T-cells
25 Tests FCIM025124
FlowCellect Mose Th17 Intracelllar Cytokine KitAnti-CD4-PerCPcloneGK1.5/Anti-IL-17-,cloneTC11-18H10-FITC
A quick and easy way to detect IL-17 expression in mouse TH17 CD4+ T-cells.
25 Tests FCIM025125
FlowCellect Mose Th1/Th2 Intracelllar Cytokine Kit20XAnti-CD4cloneGK1.5-PerCP/Anti-IL-4,clone11B11-PE/Anti-IFN-, clone XMG1.2-PE
A quick and easy way to detect IFN- and IL-4 expression in mouse Th1 and Th2 CD4+ T-cells.
25 Tests FCIM025137
FlowCellect Mose Th1/Th17 Intracelllar Cytokine KitAnti-CD4-PerCPcloneGK1.5/Anti-IL-17,cloneTC11-18H10-FITC/Anti-IFN, clone XMG1.2-PE
A quick and easy way to detect IFN- and IL-17 expression in mouse TH1 and TH17 CD4+ T-cells.
25 Tests FCIM025138
Description Qty Catalog No.
FlowCellect Mose FOXP3 Treg Identication KitAnti-CD4cloneGK1.5-PerCP/Anti-FOXP3,clone3G3-AlexaFluor647
25 Tests FCIM025126
FlowCellect Mose Viable Treg Characterization KitAnti-CD4-PerCP/Cy5.5 / Anti-CD25-PE/ Anti-Foxp3-Alexa Flour 488
25 Tests FCIM025168
Hman
Mose
Mose
Reglatory T-cell Kits
Helper T-cell Kits
3.7% 56.0%
4.2% 36.1%
FVD-660
Th-1 Cells Th-17 Cells
Side
Sc
atter 7000
9000
5000
3000
1000
0
CD4-PerCP
IFN-PE
104
104
103
103
102
102
101
101100
100
CD4-PerCP
Il-17-FITC
104
104
103
103
102
102
101
101100
100
104103102101100
FVD-660
Side
Sc
atter7000
9000
3.7% 56.0%
4.2% 36.1%
1.3% 32.5%
7.4% 58.8%
5000
3000
1000
0104103102101100
http://www.millipore.com/catalogue/item/FCIM025118?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM100158?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025123?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025124?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025125?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025137?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025138?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025126?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025168?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025168?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025126?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM100158?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025118?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025138?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025137?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025125?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025124?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025123?cid=BIOS-S-EPDF-1227-1108-DS -
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FlowCellect Hman Memory B cell
Identication Kit
We have developed a multi-parameter ow cytometry
assay or monitoring human memory B cell unction. EMD
Millipores FlowCellect Human Memory B Cell Identifcation
Kit includes three directly conjugated antibodies: Anti-HumanCD5-FITC,Anti-HumanCD19-APC,andAnti-
Human CD27-PE, along with optimized assay buers to
provide researchers the ability to phenotypically distinguish
cell types. All FlowCellect kits are optimized on guava
bench top ow cytometers. FlowCellect kits can be used on
any ow cytometer ollowing the same protocol providing
researchers a reliable and ully validated solution to study
and identiy human B cell unction right in the comorto their own lab. All three antibodies provided in the kit
are careully titrated and optimized together to ensure
maximal perormance when run in multiplex, alleviating
the need or any additional optimization.
FEATuRED PRODuCT
Isolation and Identifcation o Human Memory B cells rom human PBMCs Human memory B cells are phenotypically identifed by
using specifc human CD markers: Human CD5, CD19, and CD27. CD27 has been identifed as the key marker or identiying memory
B cells. In (A), lymphocyte populations are gated, and B cells are shown using bivariate analysis plotting CD5+CD19+ (B). Memory B
cell subsets are also identifed by using a CD19+CD27+ (C).
In healthy patients, memory B cells represent approximately 30-60% o the B-cell pool. B-cell subpopulations are deective inpatients suering rom immuno-defciency disorders, showing a reduced number o circulating CD19+CD27+ memory B cells.
Description Qty Catalog No.
FlowCellect Hman Memory B Cell Identication KitAnti-CD5cloneDK23-FITC/Anti-CD19cloneHD37-APC/Anti-CD27cloneM-T271-PE
25 test FCIM025159
FlowCellect Human B Cell FAS KitCD19-FITC,CD45-PerCPofCocktail,Anti-HumanCD95(FAS)-PE/IsotypecontrolmouseIgG1-PE
100Tests FCCH100137
Description Qty Catalog No.
FlowCellect Mose Breg Identication KitAnti-HumanCD5,clone53-7.3-APC/Anti-MouseCD19,clone1D3-FITC/Anti-MouseCD1d,clone1B1-PE/
Anti-MouseCD16/CD32,clone93Puried
25 Tests FCIM025154
Description Qty Catalog No.
FlowCellect Hman Lymphocyte ZAP-70 Characterization KitAnti-CD5-FITC/Anti-CD19-APC/Anti-ZAP-70-PE
25 Tests FCIM025122
Hman: B-cell
Mose: B-cell
Hman: T-cell Signaling
A.
Forward Scatter
Side
Sca
tter 7000
7000
9000
9000
Lymphocyte
5000
5000
3000
3000
1000
10000
0
B.
APC-CD19
FITC-C
D5
CD5+B-cell
104
104
103
103
102
102
101
101100
100
C.
APC-CD19
PE-CD27
104
104
103
103
102
102
101
101100
100
Memory B-cell
B-cell
T-cell Signaling Kit
http://www.millipore.com/catalogue/item/FCIM025159?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100137?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025154?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025122?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100137?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025122?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025154?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100137?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCIM025159?cid=BIOS-S-EPDF-1227-1108-DS -
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Immne Cell Health (Apoptosis)
FlowCellect Hman T-Cell MitoDamage Kit
EMD Millipores FlowCellect T Human T cell MitoDamage Kit
includes (1) Antibody Cocktail containing CD3-PECy5, CD4-
PE and CD8-FITC antibodies (2) MitoSense Red (1,1,3,3,3,3
-Hexamethylindodicarbocyanine iodide), a uorescent
cationic dye that accumulates in the mitochondria and is
responsive to mitochondrial potential changes and (3) 1X
Assay buer BA solution to perorm the assays. The kit
can thus distinguish multiple populations (1) CD4 T Helper
cellsand%ofthesecellswhichshowintactmitochondrial
membranepotential(2)%ofCD4THelpercellswith
dissipated mitochondrial membrane potential (3) CD8
cytotoxicTCellsandthe%ofthesecellsthatshowintact
mitochondrial membrane potential change and (4) CD8
CytotoxicTcellsand%ofthesecellsthatdemonstrate
dissipated membrane potential. The kit thus provides a
complete picture o T cell mitochondrial perturbation
status and its response or inducer treatment conditions
ordiseases.Theentireassaycanbeperformedin30min
a simple no wash manner without loss o apoptotic cells
when using PBMCs.
FEATuRED PRODuCT
Description Qty Catalog No.
FlowCellect Hman T Cell Apoptosis Kit(CD8-FITC,CD4-PE,CD3-PECy5ofcocktail/AnnexinV,CF647Reagent)
100Tests FCCH100138
FlowCellect Hman T Cell MitoDamage Kit(CD8-FITC, CD4-PE, CD3-PECy5 o cocktail/ MitoSense Red)
100Tests FCCH100139
FlowCellect Hman CD8 T Cell FAS Kit(CD8-FITC,CD3-PECY5ofCocktail/Anti-HumanCD95(FAS)-PE/IsotypecontrolmouseIgG1-PE) 100Tests FCCH100140
FlowCellect Hman T Cell Activation Kit(CD4-FITC,CD69-PE,CD3-PECY5,CD8-APCofcocktail)
100Tests FCCH100141
FlowCellect Hman CD4 T Cell FAS Kit(CD4-FITC,CD3-PECY5ofcocktail/Anti-HumanCD95(FAS)-PE/IsotypecontrolmouseIgG1-PE)
100Tests FCCH100154
FlowCellect Hman T Cell Caspase 8 Kit(CD4-PE, CD3-PECy5, CD8-APC o cocktail/ Caspase 8 FAM)
100Tests FCCH100155
FlowCellect Hman T Cell Caspase 9 Kit(CD4-PE,CD3-PECy5,CD8-APCofcocktail/Caspase9FAM)
100Tests FCCH100156
FlowCellect Hman T Cell Caspase 3/7 Kit(CD4-PE, CD3-PECy5, CD8-APC o cocktail/ Caspase 3/7 FAM)
100Tests FCCH100157
FlowCellect Hman B Cell FAS Kit(CD19-FITC,CD45-PerCPCocktail,Anti-HumanCD95(FAS)-PE/IsotypecontrolmouseIgG1-PE)
100Tests FCCH100137
Gava Cell Toxicity Kit(Guava CFSE/ CellToxicity 7-AAD)
100Tests 4500-0230
Hman
1 2
A
C
B
D
CD3-PECy5 CD4-PE
SSC
CD8
FITC
MitoSense Red
untreated Diamide
MitoSense Red
MitoSense Red
MitoSense Red
CD8+
FITC
CD4
-PE
CD8+
FITC
CD4-PE
Mitochondrial membrane
depolarization in apoptotic
CD4 and CD8 T cells
PBMCs were treated overnightwith and without 50 M
diamide and analyzed with
the FlowCellect T Cell
MitoDamage Kit. Plots show
the percentage o positive
cells or CD3, CD4 and CD8
subpopulations (1, 2) and
the mitochondrial potential
status o CD4 and CD8 T cell
populations or untreated
(A, C) and treated samples
(B, D). Cells with intact
mitochondrial potential
exhibit higher Red2 or
MitoSense red uorescence(as in untreated control),
while cells with depolarized
mitochondria demonstrate
a downward shit in their
mitochondrial potential.
http://www.millipore.com/catalogue/item/FCCH100138?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100139?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100140?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100141?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100154?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100155?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100156?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100157?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100137?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0230?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/4500-0230?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100137?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100157?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100156?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100155?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100154?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100141?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100140?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100139?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100138?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCCH100137?cid=BIOS-S-EPDF-1227-1108-DS -
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Milli-MarkConjugated Antibodies
Advance your ow cytometry analysis with EMD
Millipores growing selection o directly conjugated
primary antibodies. As a part o our complete benchtop
ow cytometry solution including instruments, sotware,
service, and reagents, Milli-Mark uorescently-labeled
antibodies are specifcally designed, optimized and
validated or ow cytometry applications.
Stem Cell
Fixed and permeabilized
2102Ep embryonal carcinomacells were stained with Milli-
Mark anti-hOct4-Alexa Fluor
488 (FCMAB113A4, green
histogram) or IgG-Alexa Fluor
488 (grey histogram) at 1:100
or 1 hour and then analyzed
by ow cytometry.
Description Reactivity Host Qty Catalog No.
Anti-hOCT4-Alexa Fluor 488 Human Mouse 100tests FCMAB113A4
Mouse
Anti-SSEA-4-PE Human Mouse 100tests FCMAB116P
Anti-Sox2-FITC Human Mouse 100tests FCMAB112F
Mouse
Rat
Anti-TRA160-FITC Human Mouse 100tests FCMAB115F
Anti-HESCA-1-FITC Human Mouse 100tests FCMAB111F
Mouse
Anti-SSEA-1-PE Human Mouse 100tests FCMAB117P
MouseRat
Anti-SSEA3Alexa488,cloneMC-631 Human Rat 100tests FCMAB141A4
Anti-Human Nuclei -FITC, clone 3E1.3 Human Mouse 100tests FCMAB157F
Anti-BMP-7-FITC,clone2A10 Human Mouse 100tests FCMAB135F
Anti-mOCT4-AlexaFluor488,clone7F9.2 Human Mouse 100tests FCMAB124A4
Mouse
Anti-TRA-1-81-FITC,clone TRA-1-81 Human Mouse 100tests FCMAB132F
Anti-TRA-2-49-FITC,cloneTRA-2-49/6E Human Mouse 100tests FCMAB133F
Anti-SRF-FITC, clone 1E1 Human Mouse 100tests FCMAB137F
Description Reactivity Host Qty Catalog No.
Anti-phospho-Bcl-2(Ser70)-PE,
clone69-10C-2-10C-18
Human Rabbit 100tests FCMAB140P
Rat-a-Caspase-2-FITC Human Rat 100tests FCMAB158F
Milli-Mark: Stem Cell
Milli-Mark: Apoptosis and Cancer
200
180
160
140
120
100
80
60
40
20
010e0 10e1 10e2 10e3 10e4
Count
hOCT4-Alexa Fluor 488
http://www.millipore.com/catalogue/item/FCMAB113A4?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB113A4?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB116P?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB112F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB115F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB111F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB117P?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB141A4?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB157F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB135F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB124A4?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB132F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB133F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB137F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB140P?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB158F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB158F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB140P?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB137F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB133F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB132F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB124A4?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB135F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB157F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB141A4?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB117P?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB111F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB115F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB112F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB116P?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB113A4?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB113A4?cid=BIOS-S-EPDF-1227-1108-DS -
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Description Reactivity Host Qty Catalog No.
Anti-MAD2A-FITC,clone17D10 Human Mouse 100tests FCMAB150F
Anti-Y14-FITC, clone 4C4 Human Mouse 100tests FCMAB151F
Anti-FXR1-FITC,clone6BG10 Human Mouse 100tests FCMAB152F
Anti-SMN-FITC, clone 2B1 Human Mouse 100tests FCMAB153F
Anti-CAF1p150-FITC,cloneSS1,1-3 Human Mouse 100tests FCMAB145FAnti-TBX21/T-Bet-FITC Human Mouse 100tests FCABS131F
Anti-RPA2p34-FITC,cloneRPA201-46 Human Mouse 100tests FCMAB143F
Anti-RPA1p70-FITC,cloneRPA9,1-30 Human Mouse 100tests FCMAB144F
Anti-BMI1-FITC, clone AF27 Human Mouse 100tests FCMAB149F
Anti-RBMS1-FITC, clone 4D11 Human Mouse 100tests FCMAB123F
Anti-c-Jun-FITC,clone6E4 Human Mouse 100tests FCMAB122F
Mouse
Rat
Anti-RBMS1-FITC, clone 4D11 Human Mouse 100tests FCMAB125F
Mouse
Anti-BrdU-Alexa 488 Human Mouse 100tests FCMAB101A4
Phospho-ATM(Ser1981)-PE Human Mouse 100tests FCMAB110P
Mouse
Rat
PhosphoH3(Ser10)-Alexa488 Human Mouse 100tests FCMAB104A4
Phospho-SMC1(Ser957)-Alexa488 Human Mouse 100tests FCMAB108A4
Bovine
Xenopus
Anti-Cyclin B1 - PE Human Mouse 100tests FCMAB102P
Mouse
Milli-Mark: Epigenetics and Gene Reglation
Epigenetics & Gene
Reglation
HeLa cells were stained with
either Milli-Mark anti-Y14-
FITC, clone 4C4 (FCMAB151F,
green histogram) or with
IgG2b isotype control antibody
(grey histogram) and analyzed
using ow cytometry.
200
180
160
140
120
100
80
60
40
20
0
10e0 10e1 10e2 10e3 10e4
Count
Green Fluorescence
http://www.millipore.com/catalogue/item/FCMAB150F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB151F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB152F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB153F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB145F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCABS131F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB143F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB144F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB149F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB123F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB122F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB125F?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB101A4?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB110P?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB104A4?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB108A4?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalogue/item/FCMAB102P?cid=BIOS-S-EPDF-1227-1108-DShttp://www.millipore.com/catalog