Cell Structure & Migration Product Selection Guide
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Transcript of Cell Structure & Migration Product Selection Guide
EMD Millipore is a division of Merck KGaA, Darmstadt, Germany
Cell Structure and MigrationAntibodies, Proteins, Inhibitors, Kits, and Assays
2
Platforms, Technologies, and ServicesAs a tools provider and partner in research, EMD Millipore is committed to the advancement of life science research and therapeutic development. This guide outlines EMD Millipore’s portfolio of small molecule libraries and pathway-focused panels, which are designed to facilitate target identification, pathway detection and profiling. These reagents provide proven solutions for a range of applications, platforms and technologies, and are backed by extensive technical support.
CALBIOCHEM® SMALL MOLECULESSmall-molecule compounds, including inhibitors,
activators, and other pathway modulators, are critical
tools for researchers studying cell signaling and other
mechanisms that control cell fate, function and phenotype.
EMD Millipore’s Calbiochem® reagents have been cited
in thousands of peer-reviewed publications. From
libraries and pathway panels to individual reagents, the
Calbiochem® line of products offers the widest and most
cited selection of inhibitors and activators worldwide.
CELLS AND CELL CULTUREEMD Millipore’s innovative cell culture solutions help
optimize cell growth and maintenance. We offer an
extensive range of human and rodent stem cells, primary
cells and media designed for most types of stem cells,
including embryonic, mesenchymal, and neural stem
cells. These optimized media include serum-free, feeder-
free formulations, and are supported by our feeder cells,
supplements, reagents, and cultureware. Our flexible sterile
filtration devices offer fast flow and have many membrane
options. Also available are membrane-based cultureware to
mimic in vivo conditions and provide coculture options.
ANTIBODIES AND IMMUNOASSAYSWith the expertise of Upstate® and Chemicon®, EMD
Millipore provides an extensive, focused, validated portfolio
of antibodies and immunoassays, with breadth and depth
in major research areas backed by excellent service and
support. EMD Millipore also offers a variety of ELISAs in
major research areas, including cell health.
CELL-BASED ASSAYS AND QUANTITATIVE CELL IMAGINGOur portfolio of live cell, whole-cell and cell-based activity
assays and reporter systems advances direct and indirect
detection of biological processes. These technologies
facilitate protein target validation, identify cellular
pathways and determine mechanism of action for lead
optimization environments. EMD Millipore also offers
assays for high-content, multiparametric cell imaging,
enabling identification of cellular responses and events
under user-defined conditions.
FLOW CYTOMETRY ASSAYS AND SYSTEMSFlow cytometry is essential for in-depth cell analysis,
with the capacity to simultaneously measure multiple
parameters on individual cells. Our easyCyte™ flow
cytometers provide direct, precise measurement via
microcapillary technology that translates into smaller
samples, less reagents, and minimal waste. Our validated
FlowCellect™ assay kits and Milli-Mark™ conjugated
antibodies, along with application-specific analysis
software modules, provide a complete solution for flow
cytometry.
MILLIPLEX® map MULTIPLEX ASSAYSMILLIPLEX® map assays offer the broadest selection of
multiplex kits and reagents in a wide variety of research
areas, measuring multiple biomarkers using a small sample
size. MILLIPLEX® map enables the simultaneous detection
of multiple soluble or intracellular biomarkers. Using the
Luminex® xMAP® bead-based technology, these flexible
and customizable assays are exhaustively tested and
qualified for sensitivity, specificity, reproducibility and wide
dynamic range.
MOLECULAR BIOLOGY TOOLSFor every step of the molecular biology and protein
workflow, from cloning DNA targets to purifying native
recombinant proteins, EMD Millipore provides reagents,
kits, cells and tools that are specifically designed to meet
your scientific and technical goals, including the Novagen®
line of products for DNA amplification, purification, and
propagation and reagents for efficient transfection.
3
Introduction
Cell Structure and MovementRecent studies elucidating the effects of extracellular
microenvironment on cell structure and movement have
revolutionized studies of diverse processes, including
stem cell differentiation, morphogenesis, wound healing,
neurogenesis, cancer metastasis, and immune response.
In just the last few years, discoveries have shown that
cell structure and movement can be characterized by
multiple, interchangeable parameters that can be adjusted
dynamically by factors such as cell type, multipotency,
signaling modulation and adhesive forces. As a result, the
study of cell structure and movement now depends on a
powerful confluence of research areas and experimental
techniques. EMD Millipore’s life scientists possess a unique
breadth and depth of research focus, as well as expertise
in developing a vast array of biochemical and visual cell
analysis methods, enabling us to offer the only complete
product portfolio for studying cell structure, intracellular
signaling, extracellular environment, and cellular migration
and invasion.
Cell structure is a broad area of research that describes
both the fundamental structural features and mechanistic
functions of a cell. A cell’s structural features, including
the cytoskeleton, filaments, microtubules and adhesion
molecules, allow the cell to perform basic mechanistic
functions such as migration, invasion, organelle transport,
cell division, extracellular matrix degradation and
restructuring, and cell-cell communication. In particular,
cell motility is an integrated process that is carefully and
precisely orchestrated by the cell with the help of many
receptors, cross linking, bundling, binding, adhesion, motor,
and other proteins. The key steps in cell movement are as
follows: (1) interaction with the environment (2) signaling,
(3) cytoskeletal rearrangement, and (4) rearrangement of
adhesive contacts to the substratum. Understanding how
the cell generates the forces necessary for movement
and learning how these forces operate at the microscopic
scale will ultimately facilitate the incorporation of similar
principles in engineering microscopic-scale tools for both
research and therapeutics.
With this future potential in mind, EMD Millipore is
continuously expanding its comprehensive portfolio of cell
structure and migration research tools to keep our customers
at the forefront of the field. This product guide represents a
sample of some of our most exciting offerings for studying
cell structure and movement.
CytoskeletonCell Adhesion Molecules
Integrins
Cell Migration
Cell Invasion
Cytoskeletal Signaling
MMPs &Proteases
Degradation
NucleusCytoskeleton-associated Protein
ECM Layer
Platforms, Technologies, and Services
4
Table of Contents
PG. 05 PG. 21
PG. 28
PG. 17
PG. 11
Migration ExtracEllular Matrix and associatEd ProtEasEs
Visualization
adhEsion
cytoskElEton and signaling
Chemotaxis• QCM™ChemotaxisMigration
Assay (fluorometric)• QCM™EndothelialMigration
Assay (fluorometric)• Millicell®μ-MigrationAssayKit
Invasion• QCM™CollagenCellInvasion
Assay (fluorometric)• Millicell®andMultiScreen-
MIC Systems
Featured Products:• QCM™GelatinInvadopodiaAssays• CellMigrationandInvasion
Product Table• PoreSizeSelectionTable
ECM• ECMCellCultureOptimizationArray• ECMProteinsandAntibodies
Proteases MMPs/TIMPs• MT1-MMPAntibody• GM6001MMPInhibitor
Featured Products:• MMP&TIMPMILLIPLEX®map Panels• MMPInhibitors• MMPProductSelectionTable• MMPActivationTable
• ConjugatedAntibodies
Technology Highlight:• LentiBrite™LentiviralBiosensors
Enrichment and Staining
• ActinCytoskeletonandFocalAdhesion Staining Kit
Featured Product:• ProteoExtract®Cytoskeleton
Enrichment and Staining Kit
Cell to ECM Interactions• α Integrin-Mediated Cell
Adhesion Colorimetric Array Kit
Cell to Cell Interactions• EndothelialCellAdhesionAssay• ECMCellAdhesionArrayKits
Featured Product:• QCM™LeukocyteTransendothelial
Cell Migration Assay
Cytoskeleton• ActinAntibodies• Acetyl-CortactinAntibody• ProteoExtract®Cytoskeleton
Enrichment and Isolation Kit
Featured Products:• AXIS™AxonInvstigationSystem• Cytoskeletonreorganization inhibitors
Signaling Proteins• FAKAntibody• RasGTPaseActivationELISAkit
5
Cell migration is stimulated and directed by interaction of cells with the extracellular matrix (ECM), neighboring cells, or chemoattractants. During embryogenesis, cell migration participates in nearly all morphogenic processes ranging from gastrulation to neural development. In the adult organism, cell migration contributes to physiological and pathological conditions, and is central to development of therapeutics affecting wound healing and tumor metastasis. Specifically,inhibitingtumorinvasionbyblockingtumorcellchemotaxishasbeenamajorfocusof research.
Migration
MMPs &Proteases
Degradation
ECM Layer
CytoskeletonCell Adhesion Molecules
Integrins
Cell Invasion
Cytoskeletal Signaling
Cytoskeleton-associated Protein
Cell MigrationCytoskeleton
Cell Adhesion Molecules
Integrins
Cell Invasion
Cytoskeletal Signaling
MMPs &Proteases
Degradation
NucleusCytoskeleton-associated Protein
ECM Layer
Cell Migration
6
The chemotaxis cell migration assay measures directional cell movement in response to
chemical concentration gradients. While multiple pore sizes are available, the 8 µm pore
size of this assay supports optimal migration for most epithelial and fibroblast cells.
see the Pore size selection chart on page 10 for more information on which pore size to choose for your specific cell type.
Angiogenesis, the formation of blood vessels, results from migration of endothelial
cells and is regulated by ECM components, angiogenic, and anti-angiogenic factors.
The endothelial cell migration assay provides a quick and efficient system to study a
compound’s ability to induce or inhibit endothelial cell migration.
QcM™ chemotaxis cell Migration assay (96-well, fluorometric) (Catalogue No. ECM510)
QcM™ Endothelial cell Migration assay (24-well, fluorometric) (Catalogue No. ECM201)
Human fibrosarcoma HT-1080 chemotaxis toward 10% fetal bovine serum (FBS) was measured in the presence or absence of cytochalasin D, an inhibitor of actin polymerization.NotethatcytochalasinDinhibitscellmigration towards chemoattractant FBS. Assayed at multiple time intervals.
Migration assay using HUVECs plated on chambers coated with bovine serum albumin (BSA) or fibronectin (FN). Fetal bovine serum functions as an effective chemoattractant, stimulating cell migration on FN but not on BSA.
Chemotaxis assays are ideal for assessing the effects of pharmacological compounds on the motility of tumor cells,andforanalyzingthemigratorycapacityofmultiplecelllinesinparallel.Themostwidelyacceptedcellmigration assay is the Boyden chamber assay. EMD Millipore’s system uses a two-chamber multiwell plate in which a membrane in each well provides a porous interface between two chambers. This user-friendly, high throughput format enables sensitive, quantitative comparison of multiple samples.
Chemotaxis
HT-1080 Chemotaxis Assay in 96-well Format
0
50
100
150
200
250
RFU
x 1
000
300
2h 4h 24h
0 % FBS
10 % FBS 10 % FBS + 10 µM Cytochalasin D
02550
RFU
x 10
00
75100125150175
BSA BSA+FBS FN+FBSFN
HUVEC Migration on Fibronectin Coated Inserts
7
The Millicell® µ-Migration Assay Kit utilizes microfluidic, low-volume technology to
promote a stable, diffusion-generated concentration gradient that is consistently linear
and lasts for more than 48 hours. Designed for video microscopy assays, the µ-Migration
Slide is made from a plastic with high optical qualities similar to those of glass. At
specific time intervals, images of the observation area can be acquired, allowing real-time
monitoring and quantitative measurements of cell migration.
Millicell® μ-Migration assay kit (Catalogue No. MMA205)
Datawasanalyzedandgraphed using a free Image J software plug-in.
description catalogue no.
QCM™ Endothelial Cell Migration Assay (24-well, colorimetric) ECM200
QCM™ Chemotaxis Assay (24-well, 3 µm, colorimetric) ECM504
QCM™ Chemotaxis Assay (24-well, 3 µm, fluorometric) ECM505
QCM™ Chemotaxis Assay (24-well, 5 µm, colorimetric) ECM506
QCM™ Chemotaxis Assay (24-well, 5 µm, fluorometric) ECM507
QCM™ Chemotaxis Assay (24-well, 8 µm, colorimetric) ECM508
QCM™ Chemotaxis Assay (24-well, 8 µm, fluorometric) ECM509
QCM™ Chemotaxis Assay (96-well, 5 µm, fluorometric) ECM512
QCM™ Chemotaxis Assay (96-well, 3µm, Fluorometric) ECM515
Fibroblast Growth Factor basic, human recombinant GF003 (multiple)
Available from www.millipore.com
Key Products
150
Cell Track0% FCS
100
50
0
y ax
is (µ
m)
x axis (µm)
Number of tracks: 78Counts up: 44Counts down: 34
Center of mass (µm): x=0.97 y=6.41
-50
-100
-100 -50 0 50 100 150-150
-150
300
Cell Track10% FCS
200
100
0
y ax
is (µ
m)
x axis (µm)
Number of tracks: 80Counts up: 51Counts down: 29
Center of mass (µm): x=0.97 y=37.25
-100
-200
-200 -100 0 100 200 300-300
-300
8
The ability to study cell invasion through a collagen barrier, is of vital importance for
developing possible metastatic inhibitors and therapeutics. The QCM™ 96-well Collagen-
based Invasion Assay provides an efficient, in vitro system for quantitative, high-
throughput analysis of tumor cell invasion.
QcM™ collagen cell invasion assay, 96-well (8 μm), fluorometric (Catalogue No. ECM556)
Millicell® inserts and Multiscreen®-Mic system The Millicell® inserts and MultiScreen®-MIC filter plates provide reliable, versatile devices
for a range of cell-based screening assays including migration, invasion, chemotaxis,
co-culture, angiogenesis, and transmigration. Inserts and plates are available in a range of
pore sizes to support assays with suspension or adherent cell lines and support cell growth
for co-culture and transmigration assays.
Thebasementmembranesurroundingthebloodvesselendotheliumisathin,specializednetworkofECMproteins. The ability of tumor cells to degrade the ECM components of the basement membrane and surroundingtissuesisdirectlycorrelatedwithmetastaticpotential.QCM™cellinvasionassaysenableconvenient and sensitive quantification of in vitro cell invasion through a basement membrane model. A Boyden chamber system is layered with an ECM solution that occludes the membrane pores, blocking non-invasive cells from migrating through it.
Invasion
description Pore size (μm) device size Qty/Pk catalogue no.
Millicell® single–well standing inserts
PCF insert Isopore (Polycarbonate)
0.4 µm 24 well 50 PIHP01250
3.0 µm 24 well PITP01250
8.0 µm 24 well PI8P01250
12.0 µm 24 well PIXP01250
0.4 µm 6 well PIHP03050
Millicell® single–well hanging inserts
PET Insert Polyethylene Terephthalate
0.4 µm 24 well 48 PIHT12R48
1.0 µm PIRP12R48
3.0 µm PISP12R48
5.0 µm PIMP12R48
8.0 µm PIEP12R48
Multiscreen®-Mic system
MultiScreen-MIC* 3.0μm 96 well 10 MAMIC3S10
5.0μm MAMIC5S10
8.0μm MAMIC8S10
* Includes a 96-well receiver plates housed in single-well trays, with lids. All parts are sterilized.
0246
RFU
x 10
00
8101214161820
No Cells(BKGD)
HT +10% FCS
HT +0% FCS
3T3+10% FCS
3T3+0% FCS
96-Well Invasion Assay (2.5 x 105 cells well with overnight invasion)
Fluorogenic detection
9
Featured Product
QcM™ gelatin invadopodia assays (green and red)(Catalogue No. ECM670 (Green Kit) and Catalogue No. ECM671 (Red Kit))
EMD Millipore’s Innovative QCM™ Gelatin Invadopodia Assays provide optimized materials
and protocols to enable reproducible analysis of invadopodia in invasive tumor cells. All of
the components necessary for affixing a thin film of pre-conjugated fluorescent matrix to
glass culture surfaces are provided. In addition, compatible reagents are provided for co-
localizing the actin cytoskeleton and nuclei with invadopodial degradation sites. This assay
may be used for assessing activity of inhibitors and promoters of invadopodia formation and
function. Finally, different cell types as well as individual cells in heterogeneous populations
may be analyzed for invasive potential with the QCM™ Gelatin Invadopodia Assay.
Features, Benefits, advantages:• Pre-conjugated Fluorescent Gelatin, available in Green and Red, saves time by eliminating
laborious conjugation steps
• Standardized protocol and optimized reagents for consistent and reproducible coating on
various glass cultureware surfaces
• Provides defined co-localization of actin-rich invadopodia and degradation sites
• Sensitive enough to visually and quantitatively measure time-courses and modulator
effects
Cellmigrationacrossafluorescently-conjugatedmatrix.Darkspots are evidence of invadopodia protrusions assisting the cell in migration. Actin cytoskeleton is stained with phalloidin (red) and overlayed with DAPI nuclear staining (blue).
description catalogue no.
QCM™ Gelatin Invadopodia Assay (Green) ECM670
QCM™ Gelatin Invadopodia Assay (Red) ECM671
QCM™ Endothelial Cell Invasion Assay (24 well, colorimetric) ECM210
QCM™ Endothelial Cell Invasion Assay (24 well, fluorometric) ECM211
QCM™ Laminin Migration Assay (24-well, colorimetric) ECM220
QCM™ ECMatrix™ Cell Invasion Assay (24-well, 8 µm, colorimetric) ECM550
QCM™ Collagen Cell Invasion Assay (24-well, 8 µm, colorimetric) ECM551
QCM™ Collagen Cell Invasion Assay (24-well, 8 µm, fluorometric) ECM552
QCM™ ECMatrix™ Cell Invasion Assay (24-well, 8 µm, fluorometric) ECM554
QCM™ ECMatrix™ Cell Invasion Assay (96-well, 8 µm, fluorometric) ECM555
QCM™ Collagen Cell Invasion Assay (96-well, 8 µm, fluorometric) ECM556
QCM™ Tumor Cell Transendothelial Migration Assay (24 well, colorimetric) ECM558
QCM™ Haptotaxis Cell Invasion Series ECM580 series
ECL Cell Attachment Matrix 08-110
TNFα Growth Factor GF023
Millicell® 24-well Receiver Tray with Lid PSMW010R5
Millicell® Single-well Receiver Tray with Lid PSSW010R5
MultiScreen® Single-well Culture Tray, clear, sterile MAMCS0110
MultiScreen® 96-well Culture Tray clear, sterile MAMCS9610
Key Products
10
cell Migration and invasion assay table description Pore size Plate Format EcM coating detection no. of tests catalogue no.
chemotaxis cell Migration assays 8 µm 24-well None Colorimetric 24 ECM50824-well Fluorometric 24 ECM50996-well Fluorometric 96 ECM510
5 µm 24-well Colorimetric 24 ECM50624-well Fluorometric 24 ECM50796-well Fluorometric 96 ECM512
3 µm 24-well Colorimetric 24 ECM50424-well Fluorometric 24 ECM50596-well Fluorometric 96 ECM515
haptotaxis cell Migration assays 8 µm 24-well Fibronectin Colorimetric 24 ECM58024-well Vitronectin Fluorometric 24 ECM58124-well Collagen I Fluorometric 24 ECM582
5 µm 24-well Laminin vials Colorimetric 24 ECM22024-well Fluorometric 24 ECM221
Millicell® μ-Migration assay kit NA 4 slides of 3 wells
NA 12 MMA205
cell invasion assays 8 µm 24-well ECMatrix™ Colorimetric 12 ECM55024-well Colorimetric 24 ECM55496-well Colorimetric 96 ECM55524-well Collagen I Colorimetric 24 ECM55124-well Colorimetric 24 ECM55296-well Fluorometric 96 ECM556
Endothelial cell Migration assays 3 µm 24-well Fibronectin Colorimetric 24 ECM20024-well Fluorometric 24 ECM201
Endothelial cell invasion assays 24-well ECMatrix™ Colorimetric 24 ECM21024-well Fluorometric 24 ECM211
leukocyte transendothelial Migration 3 µm 24-well Fibronectin Colorimetric 24 ECM557Fluorometric 24 ECM559
tumor cell transendothelial Migration
8 µm
24-well
Colorimetric 24 ECM558Fluorometric 24 ECM560
QcM™ invadopodia gelatin degradation assay (green) NA NA FITC-Gelatin* Fluorometric 32 ECM670QcM™ invadopodia gelatin degradation assay (red) Cy3-Gelatin* Fluorometric 32 ECM671
*FITC-Gelatin and Cy3-Gelatin are provided but not pre-coated.
cell name cell type Pore sizes assays typically performed
MDA-MB-231 Invasive Breast cancer cell line (human)
5 or 8 µm 5 or 8 µm used in chemotaxis or invasion assay
MCF7 Non-invasive breast cancer cell line (human)
5 or 8 µm 5 or 8 µm used in chemotaxis or invasion assay
HT1080 Invasive fibrosarcoma cell line (human)
5 or 8 µm 5 or 8 µm used in chemotaxis or invasion assay
NIH3T3 Non-invasive fibroblast cell line (mouse)
5 or 8 µm 5 or 8 µm used in chemotaxis or invasion assay
HUVEC (Human vein umbilical vein endothelial cells)
Endothelial cells 3 or 5 or 8 µm 3 or 5 or 8 µm in chemotaxis, invasion, angiogenesis or transendothelial migration assays
HMVEC/HMEC(Human dermal microvascular
Endothelial cells 5 or 8 µm 5 or 8 µm in chemotaxis, invasion, angiogenesis or transendothelial migration assays
PMN Polymorphonuclear neutrophils 1 or 3 µm 1 or 3 µm in chemotaxis assaysPrimary Stromal cells 8 µm No information availableEpithelial cells 3 or 5 µm No information availableHuman coronary artery smooth muscle cells
5μm No information available
Hepatic stellate cells Myofibroblast 5μm No information available
What pore size should i select? Pore size determination depends entirely on your cell type. A quick literature search will enable you to decide the best
pore size for the particular cells you are using. The following chart illustrates pore size choices for example cell lines used
in our assays and by some of our customers for these assays.
11
MMPs &Proteases
Degradation
ECM Layer
CytoskeletonCell Adhesion Molecules
Integrins
Cell Invasion
Cytoskeletal Signaling
Cytoskeleton-associated Protein
Cell Migration
description Pore size Plate Format EcM coating detection no. of tests catalogue no.
chemotaxis cell Migration assays 8 µm 24-well None Colorimetric 24 ECM50824-well Fluorometric 24 ECM50996-well Fluorometric 96 ECM510
5 µm 24-well Colorimetric 24 ECM50624-well Fluorometric 24 ECM50796-well Fluorometric 96 ECM512
3 µm 24-well Colorimetric 24 ECM50424-well Fluorometric 24 ECM50596-well Fluorometric 96 ECM515
haptotaxis cell Migration assays 8 µm 24-well Fibronectin Colorimetric 24 ECM58024-well Vitronectin Fluorometric 24 ECM58124-well Collagen I Fluorometric 24 ECM582
5 µm 24-well Laminin vials Colorimetric 24 ECM22024-well Fluorometric 24 ECM221
Millicell® μ-Migration assay kit NA 4 slides of 3 wells
NA 12 MMA205
cell invasion assays 8 µm 24-well ECMatrix™ Colorimetric 12 ECM55024-well Colorimetric 24 ECM55496-well Colorimetric 96 ECM55524-well Collagen I Colorimetric 24 ECM55124-well Colorimetric 24 ECM55296-well Fluorometric 96 ECM556
Endothelial cell Migration assays 3 µm 24-well Fibronectin Colorimetric 24 ECM20024-well Fluorometric 24 ECM201
Endothelial cell invasion assays 24-well ECMatrix™ Colorimetric 24 ECM21024-well Fluorometric 24 ECM211
leukocyte transendothelial Migration 3 µm 24-well Fibronectin Colorimetric 24 ECM557Fluorometric 24 ECM559
tumor cell transendothelial Migration
8 µm
24-well
Colorimetric 24 ECM558Fluorometric 24 ECM560
QcM™ invadopodia gelatin degradation assay (green) NA NA FITC-Gelatin* Fluorometric 32 ECM670QcM™ invadopodia gelatin degradation assay (red) Cy3-Gelatin* Fluorometric 32 ECM671
*FITC-Gelatin and Cy3-Gelatin are provided but not pre-coated.
CytoskeletonCell Adhesion Molecules
Integrins
Cell Invasion
Cytoskeletal Signaling
MMPs &Proteases
Degradation
NucleusCytoskeleton-associated Protein
ECM Layer
Cell Migration
Using surface receptors and adhesion molecules, cells respond to signals from other cells and from the extracellular matrix (ECM). A class of cell transmembrane proteins known as integrins mediate the adhesion of cells to ECM proteins and endothelial surfaces. These cell surface receptors anchor cells to the ECM leading to the transduction of signaling events that regulate cell survival, proliferation, and migration. These signals are translated into cell movements via tightly regulated, site-specific protein complexes that link external signals with the cytoskeleton. Recent studies have shown that transmembrane signaling is often bidirectional, allowing intracellular signals to be transmitted back to the surface proteins that control movement along extracellular scaffolds. Cytoskeletal signaling pathways control cell cycle, chromosomal organization,cellpolarity,nucleardynamics,apoptosis,tumorigenicity,andmore.
Cytoskeleton and Signaling
12
The cytoskeleton consists of actin polymers, microtubules, and intermediate filaments (IFs). Actin polymers, which mediate cellular movement and intracellular dynamics, are regulated by signaling from many proteins, including thoseoftheN-WASPfamily.Microtubules,directingmovementofsubcellularcomponents,arestabilizedbyRhoGTPases,microtubule-associatedproteins,andorganizationcenters.IFsareadiversefamilyofstructuralproteinswithequallydiversestabilizingmechanisms.Molecularmotorproteinsuseenergyfromnucleotidehydrolysistomove along actin filaments or microtubules and include myosins, kinesins, and dyneins. These proteins mediate the sliding of filaments, relative to one another, and transport organelles along cytoskeletal tracks.
Cytoskeleton
260(kDa)
1 2
160
11080
605040
30
Actin is the monomeric subunit of microfilaments, one of the three major components
of the cytoskeleton. Catalogue No. MAB1501 and its recombinant form (Catalogue No.
MAB1501R) are pan-actin antibodies that bind to a highly conserved region of actin and
recognize all six isoforms of vertabrate actin. The antibodies react with both globular (G)
and filamentous (F) forms of actin and do not interfere with actin polymerization.
Cortactin is a cytoskeleton protein that facilitates assembly of cortical actin and is a
prominent substrate of the tyrosine kinase, Src. Recent studies show cortactin is acetylated
in vivo and is deacetylated by histone deacetylase 6 (HDAC6). Cortactin acetylation regulates
actin-dependent cell motility by changing cortactin’s affinity for filamentous actin. The
acetylation level of cortactin has also been correlated with tumor metastatic potential.
EMD Millipore’s ProteoExtract® Cytoskeleton Enrichment and Isolation Kit enables
enrichment of cytoskeleton-associated proteins in as little as 20 minutes, maintains
the actin cytoskeleton and associated proteins in their native, adhered conformations,
and significantly reduces background signal, while allowing for improved biochemical
detection of low-abundance cytoskeleton-associated proteins via Western blot and mass
spectrometry.
actin, clone c4 antibodies(Catalogue Nos. MAB1501 and MAB1501R)
acetyl-cortactin antibody(Catalogue No. 09-881)
ProteoExtract® cytoskeleton Enrichment and isolation kit(Catalogue No. 17-10195)
Western Blotting analysis: Anti-acetyl-Cortactin (Catalogue No. 09-881) was used to immunoprecipitate acetylated cortactin from untreated (lane 1) or TSA/nicotinamide-treated (lane 2) HeLa cells. Immunoprecipitate was resolved by SDS-PAGE and transferred to PVDF. Blots were probed with Anti-acetyl-Cortactin. Arrow indicates acetylated cortactin (~85 kDa).
Mouse anti-actin (Catalogue No. MAB1501) staining of rat testis. Photo courtesy of Dr. Robert Wine, NIEHS.
Western Blotting analysis: Western Blot of compartmental proteins extracted from HeLa cells using ProteoExtract® Cytoskeleton Enrichment and Isolation Kit (Catalogue No. 17-10195). Immunobloting results indicate that GAPDH (Panel A) is present in the cytoplasmic and nuclear fractions (Tristan, C., et al., 2011), and the intermediate filament protein Vimentin (Panel B) is present exclusively in the cytoskeletal compartment.
250(kDa) S N
A B
C S N C
15010075
50
37
252015
13
Featured Products
axis™ axon investigation system(Catalogue Nos. AX1510, AX50010, AX45005, AX45010, AX90010)
The AXIS platform is EMD Millipore’s most advanced tool for the study of somas, neurites or
synaptic formation. A slide-mounted, microfluidic chamber system enables the deposition
and culture of neural cells and spatially controlled addition of growth factors, toxins, and
other reagents. Neurite outgrowth is restricted to narrow, parallel channels, and the resultant
outgrowth or collapse behavior is easily observed under a microscope.
Features, Benefits, advantages:• Organize, visualize, and characterize neuronal cell culture.
• Detect protein expression with better spatial resolution.
• Isolate cell bodies from axons through fluidics.
• Reduce time and expense through optimized protocols and QC validated products.
• Attain superior performance over in-house protocols.
• Optically transparent, inert, non-toxic, and nonflammable polymer mold.
• Available in 150 µm, 450 µm, 900 µm, or 6-well format.
cytoskeleton reorganization inhibitorsCytoskeletal reorganization results in dynamic structural changes to the arrangement of
constituent parts of cytoskeletons and their associated proteins, determines cell shape,
and plays important roles in diverse cellular processes such as cell division, cell motility
and migration, cellular transport, and signal transduction. Small molecule inhibitors offer
a powerful approach to identify and study the involvement of cytoskeletal reorganization
in both normal physiological and pathological processes both in cell culture in vitro and
in animals in vivo. EMD Millipore provides high quality Calbiochem® small molecules that
selectively disrupt microtubule dynamics, or potently induce, inhibit, and stabilize actin
polymerization. Calbiochem® compounds have been cited in numerous peer-reviewed
publications.
Steady-State
Volume-Mediated Hydrostatic Flow
Directional
Chemoattractant Neuronal Culture
+ Chemoattractant
description catalogue no.
Actin Polymerization Interfering Agents Set 104850
Cytochalasin D, Zygosporium mansonii 250255
Jasplakinolide, Jaspis johnstoni 420107
Latrunculin A, Latrunculia magnifica 428021
Phalloidin, Amanita phalloides 516640
Swinholide A, Theonella swinhoei 574776
Arp2/3 Complex Inhibitor I, CK-666 182515
(+)-Brefeldin A, Eupenicillium brefeldianum 203729
Colchicine, Colchicum autumnale 234115
Nocodazole 487928
Tubulin Polymerization Inhibitor 654160
Paclitaxel, Taxus sp. 580555
Order the nEW inhibitor sourcebook, third Edition by scanning this QR code:
14
description catalogue no.
Anti-DYNLT1, clone 7A9F4H10 MAB1076
Anti-DYNLT3, clone 10A3C8A1 MAB1077
Anti-Actin, clone C4 MAB1501 (multiple)
Anti-Kinesin, heavy chain, clone H2 MAB1614
Anti-Myosin X AB1094
Anti-Dynein, Left - Right AB1961
Anti-Cortactin, clone 4F11 05-180
Anti-Myosin, heavy chain, clone A4.1025 05-716
Anti-mouse Tks4 09-260
Skeletal Myogenesis Assay KAA001
ProteoExtract® Cytoskeleton Enrichment and Isolation Kit 17-10195
ProteoExtract® Cytoskeleton Enrichment and Staining Kit 17-10210
Mitochondria/Cytosol Fractionation Kit MT1000
Compartment Protein Extraction Kit 2145
ProteoExtract® Collagenase Set 71777 n
ProteoExtract® Complete Mammalian Proteome Extraction Kit 539779 n
ProteoExtract® Cytosol/Mitochondria Fractionation Kit QIA88 n
ProteoExtract® Detergent Set 539751 n
ProteoExtract® Glycopeptide Enrichment Kit 72103 n
ProteoExtract® Native Membrane Protein Extraction Kit 444810 n
ProteoExtract® Partial Mammalian Proteome Extraction Kit 539789 n
ProteoExtract® Partial Yeast Proteome Extraction Kit 539785 n
ProteoExtract® Phosphopeptide Capture Kit 525250 n
ProteoExtract® Protein Precipitation Kit 539180 n
ProteoExtract® S-PEK Antibody Control Kit 71771 n
ProteoExtract® Subcellular Proteome Extraction Kit 539790 n
ProteoExtract® Subcellular Proteome Extraction Kit, Mini 539791 n
ProteoExtract® Tissue Dissociation Buffer Kit 539720 n
ProteoExtract® Transmembrane Protein Extraction Kit 71772 n
Key Products
n Order at www.emdbiosciences.comAll others can be ordered at www.millipore.com
15
CytoskeletalsignalingcomplexesincludeG-proteincomplexes,focaladhesions,andadherensjunctions.TheRhofamily of small G-proteins transmit mechanical signals from the plasma membrane. Rho family members Rac, Rho and Cdc42 regulate the assembly of actin-based lamellipodia, stress fibers and filopodia, respectively. They also mediate polarity, proliferation, apoptosis, membrane transport, and gene expression. Focal adhesions and adherens junctionsaremembrane-associatedcomplexesthatlinkthecellexteriortotheplasmamembraneandtheactincytoskeleton. Focal adhesions are key initiators of signaling in response to adhesion.
Signaling Proteins
Each Pathway Explorer Antibody Minipack contains three related antibodies as part of a
signaling cascade or a combination of total and phosphorylated forms of key signaling
targets. Each of the three antibodies are 30% the original pack size. The Focal Adhesion
Pathway Explorer Antibody Minipack contains smaller sizes of Anti-FAK, clone 4.47
(Catalogue No. 05-537), Anti-Src, clone GD11 ((Catalogue No. 05-184) and Anti-phospho-
Paxillin (Tyr118) (Catalogue No. 07-733). Full size versions of each of the Pathway Explorer
antibodies are available for sale individually.
Ras GTPases regulate cell growth, proliferation, differentiation, and survival. Inactive Ras
is bound to GDP. Active, GTP-bound Ras changes conformation, allowing it to bind to
effectors, including members of the Raf family, the RalGDS family and PI3-kinase. The Ras
activation ELISA kit detects activated Ras using a chemiluminescent substrate.
Focal adhesion Pathway Explorer antibody MiniPack (Catalogue No. 15-113)
ras gtPase activation Elisa kit (96-well) (Catalogue No. 17-424)
Western Blot Analysis:Non-stimulated A431 cell lysate was probed with anti-Src(0.5μg/mL).ArrowindicatesSrc(~60kDa).
Ras GTPase Activation ELISA Kit (Catalogue No. 17-424). In lysates of EGF-treated HeLa cells, levels of activated Ras are elevated as compared to lysates of untreated HeLa cells.
97
66
45
31
21
0
20
Activ
atio
n (R
LU x
100
0)
40
60
80
100
120
140
160
0 25
HeLa Cell Lysate (µg/well)25
Assay Background
Unstimulated HeLa Cell Lysate
EGF Stimulated HeLa Cell Lysate
RasGTPase Activity Assay
16
description catalogue no.
Focal Adhesion Pathway Explorer Antibody MiniPack 15-113
Anti-Rho (-A, -B, -C), clone 3L74 04-822
Anti-Rac1, clone 23A8 05-389
Anti-Cdc42 07-1466
description catalogue no.
FAK STAR ELISA 17-479
Rac1/Cdc42 Activation Assay Kit 14-441
Ras Activation ELISA Assay Kit 17-497
MILLIPLEX® map 8-Plex Human Src Family Kinase Kit – Phosphoprotein 48-650
Rho Activation Assay Kit 17-294
Rac1 Activation Assay Kit 17-283
Cdc42 Activation Assay Kit 17-286
description catalogue no.
Rho/SRF Pathway Inhibitor, CCG-1423 555558-25MG
Rac1 Inhibitor 553502-10MG
Toxin A, Clostridium difficile 616379-2UG
Key Products
antibodies
assays
inhibitors
17
MMPs &Proteases
Degradation
ECM Layer
CytoskeletonCell Adhesion Molecules
Integrins
Cell Invasion
Cytoskeletal Signaling
Cytoskeleton-associated Protein
Cell Migration
Cell to cell and cell to extracellular matrix (ECM) adhesion enables cell communication and movement, and is critical for the development and maintenance of tissues. Adhesion is involved in immune cell migration, metastasis of tumor cells, angiogenesis, wound healing, and other processes. Changes in cell adhesion can be the defining event in a wide range of diseases, including cancer, osteoporosis, atherosclerosis, and arthritis. Adhesion between tumor cells and their microenvironment involves integrins, matrix metalloproteases (MMPs), and the ECM, and is of particular interest as a targeted pathway for cancer therapy.
Adhesion
Cytoskeleton
Cell Invasion
Cytoskeletal Signaling
MMPs &Proteases
Degradation
NucleusCytoskeleton-associated Protein
ECM Layer
Cell Migration Cell Adhesion Molecules
Integrins
18
Adhesion molecules that interact with the ECM enable the cell to move or maintain position. The complex network of proteins and proteoglycans that make up the ECM can interact simultaneously with multiple cell surface receptors. The most abundant ECM proteins are fibronectin, laminin, and collagen. Many interactions with the ECM are mediated by integrins, which transduce bidirectional signals controlling diverse processes, including cell growth, differentiation, migration, and apoptosis.
Cell to ECM Interactions
The majority of cell-ECM interactions occur via integrins, a large family of heterodimeric
membrane glycoproteins consisting of noncovalently associated α and b subunits.
Integrin combinations can either recognize a single ECM ligand or bind several different
ECM proteins. This colorimetric integrin array kit is a cost effective, efficient method for
identification of surface integrins, and is a more rapid and quantitative than traditional
methods.
α integrin-Mediated cell adhesion array kit (colorimetric) (Catalogue No. ECM530)
α integrin adhesion Profile of Various cell lines. Several cell lines were incubated with an array of individually-coated α-integrin antibodies ona96-wellplate.Thegraph(right) shows differing integrin expression levels for each cell line. Adding or altering experimental factors to this type of test system allows one to monitor the effects on the adhesion signaling.
description catalogue no.
Anti-Heparan Sulfate Proteoglycan, clone 7E12 MAB458
Anti-Heparan Sulfate Proteoglycan (Perlecan), clone 5D7 MABT12
Anti-Integrin α6, clone NK1-GoH3 MAB1378
Anti-Integrin αVb3, clone LM609 MAB1976 (multiple)
Anti-Laminin-g2, clone D4B5 MAB19562 (multiple)
Anti-Glypican-1, clone 4D1 MAB2600
Anti-Syndecan-1 AB8230
Other Integrin-Mediated Cell Adhesion Arrays ECM530 series
Integrin αVb3 purified protein CC1019
Integrin αVb5 purified protein CC1025
Key Products
OD a
t 560
nm
1.8
HL-60HT-1080KU812RBLU937
1.6
1.4
1.2
1.0
0.8
0.6
0.4
0.2
0.0α1 α2 α3 α4 α5 αV αV/β3 Negative
α Integrin Antibodies
19
Illustrated by tissue patterning in multicellular organisms, the role of cell-cell adhesion molecules is often to controlmigration,trafficking,organization,andfinalanchoringofspecificcelltypestotheirniche.Majorfamiliesof adhesion molecules include ephrins, cadherins (calcium dependent), immunoglobulin-like adhesion molecules (CAMS), and selectins (carbohydrate binding). Cell-cell adhesion molecules form boundaries and gradients that maintain tissue integrity in multicellular organisms and facilitate cell-cell communication.
Cell to Cell Interactions
Both extravasation and intravasation are crucial steps during tumor cell metastasis, and
both processes require tumor cell adhesion to the endothelial cells of blood vessel walls. The
Endothelial Cell Adhesion Assay measures the affinity between circulating tumor cells and
endothelial cells in response to manipulation of activated or inactivated endothelial cells.
advantages include:• Simple fluorometric screening format
• Endothelial activator and inhibitor controls
• Blocking antibodies included
Examine the adhesion, migration, and invasion of many diverse cell types on various ECM
proteins with the ECM Cell Adhesion Array Kits. The kits provide an ECM array microtiter
plate with a homogenous detection format, allowing for large-scale screening and
quantitative comparison of multiple samples or cell lines. The kit is designed as a rapid
(1-2 hours), efficient method for characterization of cell adhesion.
Endothelial cell adhesion assay (Catalogue No. ECM645)
EcM cell adhesion array kits (Catalogue Nos. ECM540 (colorimetric) and ECM545 (fluorometric))
adhesion of hl60 and thP-1 cells to huVEcs measured by Endothelial adhesion assay kit (EcM645). Human Vascular Endothelial cells (HUVECs) were cultured and treated with or without cycloheximide. The cells were then activated with TNFα or IL-1b.Cycloheximideblocked>90%ofHL60andTHP-1celladhesion to the endothelium.
ECM Protein Array Cell Adhesion Profile
0
10
20
30
40
50
RFUs
(485
/530
nm
x 1
000) 60
Collagen I Collagen II Collagen IV Fibronectin LamininECM Proteins
Tenascin Vitronectin Blank
HBK293
HT1080
NH3T3
CHO
MC3T3 E1
HUVECs
EcM Protein cell adhesion Fluorometric array. The ECM adherence profile of several cell lines is displayed across various ECM proteins. Note that each cell line has a unique adhesion profile.
0
2
4
6
8
10
12
14
(+)Cycloheximide(+) Cytokine
(+)Cycloheximide(-) Cytokine
(-)
Cycloheximide(+) Cytokine
(-)Cycloheximide(-) Cytokine
HL60 IL-1βTreatment
HL60 TNFαTreatment
THP-1 IL-1βTreatment
THP-1 TNFαTreatment
Adhesion Assay Conditions
Cyclohexamide Blocks Adhesion to Endothelial Cells
RFUs
x 1
04
ECM Protein Array Cell Adhesion Profile
0
10
20
30
40
50
RFUs
(485
/530
nm
x 1
000) 60
Collagen I Collagen II Collagen IV Fibronectin LamininECM Proteins
Tenascin Vitronectin Blank
HBK293
HT1080
NH3T3
CHO
MC3T3 E1
HUVECs
ECM Protein Array Cell Adhesion Profile
0
10
20
30
40
50
RFUs
(485
/530
nm
x 1
000) 60
Collagen I Collagen II Collagen IV Fibronectin LamininECM Proteins
Tenascin Vitronectin Blank
HBK293
HT1080
NH3T3
CHO
MC3T3 E1
HUVECs
20
B.
a.
Featured Product
QcM™ leukocyte transendothelial cell Migration assay(Catalogue No. ECM559)
In order for leukocytes to migrate to distant locations of infection or inflammation, they
must circulate in the bloodstream, then migrate out of a blood vessel (extravasation). The
penetration of circulating leukocytes into the endothelium is a crucial step in the process.
This transendothelial cell migration assay enables quantitative analysis of the ability of
leukocytes to invade the endothelium.
Features, Benefits, advantages:• Statistical relevant analysis of multiple samples
• Consistent, convenient solution to a complex assay
• Fluorescent labeling technique returns highly specific results
description catalogue no.
Anti-PECAM1, azide-free, clone 2H8 MAB1398Z
Anti-VE-cadherin, clone BV6 MAB1989
Anti-E-selectin, clone P2H3 MAB2150
Anti-Connexin 43, C-terminus, clone 4E6.2 MAB3067
Anti-E-cadherin, azide-free, clone 67A4 MAB3199Z
Anti-MCAM (CD146), azide-free, clone P1H12 MAB16985
Anti-Connexin 45, near C-terminus, cytoplasmic AB1745
E-selectin ELISA Kit ECM330
ICAM-1 ELISA Kit ECM335
VCAM ELISA Kit ECM340
QCM™ Leukocyte Transendothelial Migration Assay (24 assays, colorimetric) ECM557
QCM™ Tumor Cell Transendothelial Migration Assay (24 assays, fluorometric) ECM560
transendothelial Migration of leukocytes. Migration of HL-60cellsthroughstimulated(TNFα) and unstimulated endothelialcelllayerover6hours,usingfetalbovineserum (FBS) as a chemoattractant. The data in (A) indicates that a greater number of leukocytes invade an activated endothelial cell system over unstimulated conditions. Note, however, over an extended period of time(18hours),HL-60cellseventuallymigratetowardsan FBS chemoattractant as outlined in (B). Additionally, an inactive endothelial layer does not provide the appropriate signaling cascade for intravasation of leukocytes. HUVECs were grown to confluency on cell culture inserts provided.
Key Products
1
2
4
3
0
10%
FBS
RFU
x 10
00
10%
FBS
321
4
98765
0
RFU
x 10
00
ControlTNFα
Control10% FBS
21
MMPs &Proteases
Degradation
ECM Layer
CytoskeletonCell Adhesion Molecules
Integrins
Cell Invasion
Cytoskeletal Signaling
Cytoskeleton-associated Protein
Cell Migration
The extracellular matrix (ECM) surrounds and supports the cells within living systems. The most prominent class of ECM proteins consists of structural proteins of the collagen and elastin families.Collagenfibersstrengthenandorganizethematrix;elastinfibersprovideflexibilityandresilience. Adhesive proteins such as fibronectin, laminin, and tenascin enable cell attachment and form cross-links within the matrix. In addition, numerous proteoglycans and heparan sulfate-containingproteinsformthehydrated,gel-likemixturethathelpsstabilizethematrixwithin its aqueous environment.
Extracellular Matrix and Associated Proteases
MMPs &Proteases
Degradation
ECM Layer
CytoskeletonCell Adhesion Molecules
Integrins
Cell Invasion
Cytoskeletal Signaling
NucleusCytoskeleton-associated Protein
Cell Migration
22
Extracellular matrix (ECM) proteins are produced intracellularly and are subsequently secreted into the surrounding cellular medium, actively regulating a diverse range of cell functions including cell adhesion, differentiation, proliferation, migration, invasion, and survival.
ECM Proteins
Adding ECM proteins to in vitro cell cultures can promote cellular adhesion while
maintaining cell viability and maximizing cell proliferation for downstream cell-based
applications. The ECM Cell Culture Optimization Array provides a multiparametric assay
that quickly identifies ECM protein(s) and pinpoints optimal concetrations that best
promote cell-type specific adhesion for a particular cell type.
Different cell types have different affinities for various ECM proteins, depending on which
cell surface proteins are expressed. EMD Millipore offers a wide variety of ECM proteins
and antibodies to meet the Individual growth needs of any cell line.
To search for your ECM target, visit www.millipore.com
EcM cell culture optimization arrays (96-well, fluorometric) (Catalogue No. ECM546)
EcM Proteins and antibodies
rencell® VM adhesion profiling against EcMs. Human neural stem cells, ReNcel®l VM, were seeded on the ECM Cell Culture OptimizationArrayat105cellsperwellfor2hoursat37°C.Cell adhesion levels were measured by crystal violet staining and analyzedbyspectrometer.Eachdatasetrepresentsthreereplicates.
Mouse anti-Fibronectin, cs-1. Staining of normal human gut mucosa. (Cat. No. MAB1939)
description catalogue no.
Quantimatrix® Human Fibronectin ELISA ECM300
Quantimatrix® Human Laminin ELISA ECM310
3D Collagen Culture Kit ECM675
Osteogenesis Assay ECM810
In Vitro Osteogenesis Quantitative Assay ECM815
Collagen Type I, rat tail 08-115
Mouse Laminin CC095
Proteoglycan CC117
Human Plasma Fibronectin Purified Protein FC010 (multiple)
Soluble RANK Ligand (sRANKL), recombinant human GF091
Wnt-3a, recombinant mouse GF160
Mouse Bone Metabolism Panel 1A MBN1A-41K
Rat Bone Metabolism Panel 1 RBN1-31K
Mesenchymal Stem Cell Osteogenesis Kit SCR028
Key Products
0.0
0.5
1.0
1.5
Optic
al D
ensi
ty
(OD
570
nm)
ECM (µg/mL)
ReNcell VM
0 5 10 15 20 25
LN
FN
COL
VN
23
Matrix metalloproteinases (MMPs) are secreted or transmembrane proteolytic endopeptidases thatprocess and degrade extracellular matrix proteins. MMPs play critical roles in many normal growth and developmental aspects of tissue remodeling, wound healing, and angiogenesis. In a pathological context, MMPs are associated with cell migration,invasion,arthritis,andcancertumorprogression.Onceactivated,theMMPsaresubjecttoinhibitionby the tissue inhibitors of metalloproteinases (TIMPs) that bind MMPs non-covalently. Included in this group are the ADAM proteins. The name ADAM stands for “A Disintegrin And Metalloprotease” and like the name suggests, ADAM proteins are cell surface proteins that possess both a adhesion domain as well as a protease domain.
Metalloproteinases
Most MMPs are secreted as inactive proproteins which are activated when cleaved by
extracellular proteases. However, MT1-MMP (MMP-14) is a member of the membrane-type
subfamily. Each member of this subfamily contains a potential transmembrane domain
suggesting that they are expressed at the cell surface rather than secreted.
MT1-MMP mediates pericellular proteolysis of ECM components and is important for
matrix remodeling. This protein also activates MMP2 protein, and this activity may be
involved in tumor invasion.
GM6001 (Ilomastat or Galardin) is a potent broad-spectrum hydroxamate inhibitor of
matrix metalloproteinases (MMPs) that can be used in both in vitro and in vivo applications
to assess the biological effects of perturbing MMP activity.
Mt1-MMP antibody (Catalogue No. AB6004)
gM6001 MMP inhibitor (Catalogue Nos. CC1000, CC1010, CC1100)
Immunohistochemcial staining of MT1-MMP in paraffin-embedded colorectal carcinoma tissue using anti-MT1-MMP, hingeregion(CatalogueNo.AB6004).Patterndepictsacytoplasmic to diffuse membrane staining of malignant cells.
GM6001inhibitsMMP-2activityatconcentrationsgreaterthan 0.1 nM.
010
MM
P-2
Activ
ity (%
)
2030405060708090
100
0 0.01 0.1 1.0 10 100GM6001 (nM)
Dose-Response for GM6001 effect on MMP-2 Activity
description catalogue no.
Quantimatrix® Human Fibronectin ELISA ECM300
Quantimatrix® Human Laminin ELISA ECM310
3D Collagen Culture Kit ECM675
Osteogenesis Assay ECM810
In Vitro Osteogenesis Quantitative Assay ECM815
Collagen Type I, rat tail 08-115
Mouse Laminin CC095
Proteoglycan CC117
Human Plasma Fibronectin Purified Protein FC010 (multiple)
Soluble RANK Ligand (sRANKL), recombinant human GF091
Wnt-3a, recombinant mouse GF160
Mouse Bone Metabolism Panel 1A MBN1A-41K
Rat Bone Metabolism Panel 1 RBN1-31K
Mesenchymal Stem Cell Osteogenesis Kit SCR028
24
Featured Product
MMP and tiMP MilliPlEx® map Panels(Catalogue Nos. HMMP1-55K, HMMP2-55K, HTIMP1-54K, HTIMP2-54K)
Based on the Luminex® xMAP® multiplex platform, MILLIPLEX® map Human MMP Panel 1
(MMP-3, -12, -13) and MMP Panel 2 (MMP-1, -2, -7, -9, -10) will enable you to explore
modulation and function of MMP expression by complete profiling of multiple MMP
expression in a single sample.
The MILLIPLEX® map Human TIMP Panels enable you to explore the modulation of TIMP
expression and the function of the MMP/TIMP balance in a variety of therapeutic areas.
Human TIMP Panel 1 (TIMP-1, -2) is to be used for serum/plasma samples, while Human TIMP
Panel 2 (TIMP-1, -2, -3, -4) has been designed for tissue/cell culture samples. Both panels are
customizable, allowing you to choose the analytes that best fit your needs.
description catalogue no.
MILLIPLEX® map Human MMP Panel 1 HMMP1-55K
MILLIPLEX® map Human MMP Panel 2 HMMP2-55K
MILLIPLEX® TIMP Human MMP Panel 1 HTIMP1-54K
MILLIPLEX® TIMP Human MMP Panel 2 HTIMP2-54K
description catalogue no.
Anti-MT1-MMP, clone 3G4.2 MAB1767
Anti-MT1-MMP AB8345
Anti-TIMP-3, C-terminus AB6000
Anti-MMP-1, hinge region AB6002
Anti-MMP-2, hinge region AB6003
Anti-TIMP-1, C-terminus AB6007
Anti-MMP-9 AB19016
Anti-MMP-1, clone 4E7.2 MAB13439
Anti-MMP-2, hinge region AB6003
Anti-MMP-3, hinge region AB810
Anti-MMP-7, catalytic domain AB8117
Key Products
25
description catalogue no.
MILLIPLEX® map Human MMP Panel 1 HMMP1-55K
MILLIPLEX® map Human MMP Panel 2 HMMP2-55K
MILLIPLEX® TIMP Human MMP Panel 1 HTIMP1-54K
MILLIPLEX® TIMP Human MMP Panel 2 HTIMP2-54K
description catalogue no.
Anti-MT1-MMP, clone 3G4.2 MAB1767
Anti-MT1-MMP AB8345
Anti-TIMP-3, C-terminus AB6000
Anti-MMP-1, hinge region AB6002
Anti-MMP-2, hinge region AB6003
Anti-TIMP-1, C-terminus AB6007
Anti-MMP-9 AB19016
Anti-MMP-1, clone 4E7.2 MAB13439
Anti-MMP-2, hinge region AB6003
Anti-MMP-3, hinge region AB810
Anti-MMP-7, catalytic domain AB8117
Featured Product
EMd Millipore’s MMP inhibitorsMatrix Metalloproteinases (MMP) are a family of zinc metallo-endopeptidases secreted by
normal and tumor cells and are responsible for much of the turnover of extracellular matrix
components. Normal physiological roles for MMPs include neurite growth, cell migration,
bone elongation, wound healing and angiogenesis. Pathological processes involving MMPs
include tumor growth and migration, fibrosis and arthritis.
Small molecule inhibitors offer a powerful approach to identify and study the involvement
of MMPs in both normal physiological and pathological processes both in cell culture in vitro
and in animals in vivo. Calbiochem® high quality small molecules potently inhibit a broad-
spectrum of MMPs and have been cited in numerous peer-reviewed publications.
description catalogue no.
GM6001 MMP Inhibitor CC1000, CC1010, CC1100
MMP-9 Inhibitor I 444278
MMP-2/MMP-9 Inhibitor II 444249
MMP-9/MMP-13 Inhibitor I 444252
MMP Inhibitor I 444250
MMP-2/MMP-9 Inhibitor I 444241
MMP-2 Inhibitor III 444288
MMP-2 Inhibitor I 444244
MMP-2/MMP-9 Inhibitor III 444251
targetinhibitor catalogue no. inhibitor
antibody catalogue no. antibody*
Protein catalogue no. Protein*
MMP-1 / MMP-8
CC1000, CC1010, CC1100
GM6001 Broad Spectrum MMP Inhibitor
MAB3307 Anti-MMP-1, clone 41-1E5 444208 MMP-1, Proenzyme, Hu Rheumatoid Synovial Fibroblast
444250 MMP Inhibitor I AB8105 Anti-MMP-1, C-terminus 444229 MMP-8, Hu NeutrophilMAB3316 Anti-MMP-8, clone 115-13D2AB81016 Anti-MMP-8, N-terminus
MMP-2 / MMP-9
444288 MMP-2 Inhibitor III MAB3308 Anti-MMP-2, clone 42-5D11 PF023-5UG MMP-2, Active, Hu Recombinant, CHO Cells
444244 MMP-2 Inhibitor I AB19167 Anti-MMP-2, whole molecule 444213 MMP-2, Proenzyme, Hu Rheumatoid Synovial Fibroblast
444249 MMP-2/MMP-9 Inhibitor II MAB13405 Anti-MMP-2 Proform, N-terminus, clone CA-4001
PF037-10UG MMP-2, Proenzyme, Hu Recombinant, CHO Cells
444241 MMP-2/MMP-9 Inhibitor I AB19016 Anti-MMP-9, Catalytic domain CC079 MMP-9, Hu transfected cells (pro and active)
444251 MMP-2/MMP-9 Inhibitor III AB13458 Anti-MMP-9, clone 9D4.2 CC1048 MMP-9, Hu, proform444278 MMP-9 Inhibitor I MAB13415 Anti-MMP-9, clone GE-213 PF038-10UG MP-9, Proenzyme, Hu
Recombinant, CHO CellsMMP-9 / MMP-13
444252 MMP-9/MMP-13 Inhibitor I AB8120 Anti-MMP-13 444287 MMP-13 Hu Recombinant, ActiveMAB13426 Anti-MMP-13, clone LIPCO IID1 CC068 MMP-13, Human, truncated,
recombinant
MMP Product selection table This is just a sample list of products available from EMD Millipore. Search our website for our entire line of MMP antibodies, inhibitors and enzymes!
Collagenase Family MMP-1, 8, 13, 18
Signal/Prodomain Catalytic Prodomain Hinge Region
Hemopexin
Gelatinase FamilyMMP-2, 9
Stromelysin FamilyMMP-3, 10, 11
Member-Type FamilyMMP-14, 15, 16, 17
Matrilysin MMP-7
PRCGVPD Gelatin Binding
Transmembrane
AXKR
26
Enzyme synonymslatent Form MWappr (kda) activating agent conditions
activation termination
active Form MWappr (kda) Mature active Form n-terminal
sequence
relativeactivation
level referenceintermediates Mature Form calculated
MMP-1 Collagenase-I, Fibroblast Collagenase, Intersitital Collagenase
57Gly and 52aa=51.8acGly~1x5
APMA, 1mM 37˚C, 2-4h A 44 43 42.5 V82LTEG and L83TEGN
1x Grant et al., jbc 1987, 262, 5886; Fields et al. Biochemistry 1990, 29, 6671; Windsor et al., JBC 1994, 269,26201; Goldberg et al., JBC 1986, 261, 6600
TrypsinTPCK, 10 µg/mL 37˚C, 10-20 min B 47
active-MMP-7 (1:1 molar ratio)
37˚C, slow C 43 42.6 F81VLTEG low Imai et al., jbc270/1995/6691; Reinemer et al., FEBS Lett. 1994/338/227
APMA 1 mM + MMP-7 (1:1)
37˚C, 2-6h 6.5x
active-MMP-3 37˚C; slow 43 42.6 F81VLTEG low Suzuki et al., Biochem 1990/29/10261, Nagase et al. Matrix Suppl. 1992 1:237, Murphy et al., Biochem J. 1987 248, 265
APMA 1 mM + active-MMP-3
37˚C 41C-term 40.8 5-8x
MMP-2 Gelatinase A, 72kDa Gelatinase, Type IV Collagenase
72 aa=71.0
APMA, 1mM 37˚C, 1-2h A 62 62.1 Y81NFFPR 1x Stetler-Stevenson et al., jbc 1989, 264, 1353; Nagase et al., Matrix Suppl. 1992 1:237 Crabbe FEBS Lett 1994 345 14, Okada Eur j biochem 1990 194 721
TrypsinTPCK, 10 µg/mL does not activate
active-MMP-3 does not activate
active-MMP-7 37˚C, 8h C n.d. 0.6xMMP-3 Stromelysin-1 (59 & 57)Gly
aa=52.2acGly~1x5
APMA, 1 mM 37˚C, 6-12h A 46 47Gly and 45 42.8 F83RTFPG 1x Galazka bioch 1996 1996/34/11221; Nagase,bioch 1990/29/ 5783; Chen et al Biochem 1993/39/10289 Docherty, Murphy Ann Rheum.Dis 1990/49/469 Harrison, etal. Biochem 31,1992, 10757
TrypsinTPCK, 10 µg/mL 37˚C, 30 min B 53 47Gly and 45 42.8 F83RTFPG 1xactive-MMP-7 does not
activate
MMP-7 Matrilysin, PUMP 28 aa=27.9
APMA, 1 mM 37˚C, 1h A 19 19.1 Y78SLFPNS 1x Imai et al., JBC270/1995/ 6691 Crabbe, et.al. Biochem. 31,1992,8500
TrypsinTPCK, 10 µg/mL 37˚C, 1-2h BMMP-3 (5x molar excess)
37˚C, 8-16h C
MMP-8 Collagenase-2, Neutrophil collagenase
(75 & 58N-trm)Gly
aa=51.1 ppGly~2x5 acGly~3x5
APMA, 1 mM 37˚C, 1-3h A 58Gly 41.9 M80LTPGNP and L81TPGNP
1x Mallya Biochem 1990/29/10628; Tschesche Matrix Suppl. 1992 1/245
TrypsinTPCK, 10 µg/mL 37˚C, slow B SlowMMP-3 C 58Gly 41.9 F79MLTPGNP Superact. Knauper, Biochem J.
1993/295/581; Reinemer, FEBS Lett. 1994/338/227; Farr et al Biochem. 1999; 38, 7332
MMP activation tableMatrix Metalloproteinases (MMPs) are synthesized as latent pro-enzymes that require activation. The following table provides activating agents, termination conditions and relative activation levels designed as a
resource & reference guide for activating various MMPs.
27
Enzyme synonymslatent Form MWappr (kda) activating agent conditions
activation termination
active Form MWappr (kda) Mature active Form n-terminal
sequence
relativeactivation
level referenceintermediates Mature Form calculated
MMP-1 Collagenase-I, Fibroblast Collagenase, Intersitital Collagenase
57Gly and 52aa=51.8acGly~1x5
APMA, 1mM 37˚C, 2-4h A 44 43 42.5 V82LTEG and L83TEGN
1x Grant et al., jbc 1987, 262, 5886; Fields et al. Biochemistry 1990, 29, 6671; Windsor et al., JBC 1994, 269,26201; Goldberg et al., JBC 1986, 261, 6600
TrypsinTPCK, 10 µg/mL 37˚C, 10-20 min B 47
active-MMP-7 (1:1 molar ratio)
37˚C, slow C 43 42.6 F81VLTEG low Imai et al., jbc270/1995/6691; Reinemer et al., FEBS Lett. 1994/338/227
APMA 1 mM + MMP-7 (1:1)
37˚C, 2-6h 6.5x
active-MMP-3 37˚C; slow 43 42.6 F81VLTEG low Suzuki et al., Biochem 1990/29/10261, Nagase et al. Matrix Suppl. 1992 1:237, Murphy et al., Biochem J. 1987 248, 265
APMA 1 mM + active-MMP-3
37˚C 41C-term 40.8 5-8x
MMP-2 Gelatinase A, 72kDa Gelatinase, Type IV Collagenase
72 aa=71.0
APMA, 1mM 37˚C, 1-2h A 62 62.1 Y81NFFPR 1x Stetler-Stevenson et al., jbc 1989, 264, 1353; Nagase et al., Matrix Suppl. 1992 1:237 Crabbe FEBS Lett 1994 345 14, Okada Eur j biochem 1990 194 721
TrypsinTPCK, 10 µg/mL does not activate
active-MMP-3 does not activate
active-MMP-7 37˚C, 8h C n.d. 0.6xMMP-3 Stromelysin-1 (59 & 57)Gly
aa=52.2acGly~1x5
APMA, 1 mM 37˚C, 6-12h A 46 47Gly and 45 42.8 F83RTFPG 1x Galazka bioch 1996 1996/34/11221; Nagase,bioch 1990/29/ 5783; Chen et al Biochem 1993/39/10289 Docherty, Murphy Ann Rheum.Dis 1990/49/469 Harrison, etal. Biochem 31,1992, 10757
TrypsinTPCK, 10 µg/mL 37˚C, 30 min B 53 47Gly and 45 42.8 F83RTFPG 1xactive-MMP-7 does not
activate
MMP-7 Matrilysin, PUMP 28 aa=27.9
APMA, 1 mM 37˚C, 1h A 19 19.1 Y78SLFPNS 1x Imai et al., JBC270/1995/ 6691 Crabbe, et.al. Biochem. 31,1992,8500
TrypsinTPCK, 10 µg/mL 37˚C, 1-2h BMMP-3 (5x molar excess)
37˚C, 8-16h C
MMP-8 Collagenase-2, Neutrophil collagenase
(75 & 58N-trm)Gly
aa=51.1 ppGly~2x5 acGly~3x5
APMA, 1 mM 37˚C, 1-3h A 58Gly 41.9 M80LTPGNP and L81TPGNP
1x Mallya Biochem 1990/29/10628; Tschesche Matrix Suppl. 1992 1/245
TrypsinTPCK, 10 µg/mL 37˚C, slow B SlowMMP-3 C 58Gly 41.9 F79MLTPGNP Superact. Knauper, Biochem J.
1993/295/581; Reinemer, FEBS Lett. 1994/338/227; Farr et al Biochem. 1999; 38, 7332
Enzyme synonymslatent Form MWappr (kda) activating agent conditions
activation termination
active Form MWappr (kda) Mature active Form n-terminal
sequence
relativeactivation
level referenceintermediates Mature Form calculated
MMP-9 92kDa Gelatinase, Gelatinase B
92Gly aa=76.3 ppGly~1x5 acGly~2x5
APMA, 1mM 37˚C, 16-24h A 83 67 66.6* M75RTPR 1x Wilhelm et al., 1989, Imai,JBC270/1995/6691; Okada, JBC267,1992, 21712; Shapiro JBC 270,1995,6351
TrypsinTPCK, 10 µg/mL 37˚C, 2h B 74 and 68 64 66.6* A74MRTPR; C-term cleavage
1x Tschesche et al.1992, Matrix Supp 1. 245, Okada, JBC267/1992/21712
activeMMP-7 1:1 37˚C, 4h C 83 & 80 78 74.6* L16RTNL; C-term cleavage
0.25x Imai, jbc270/1995/6691
APMA, 1 mM + activeMMP-7 (1:1)
37˚C, 12h 83 62 66.6* M75RTPR & F88QTFE; C-term
cleavage
0.7x Imai, jbc270/1995/6691
Active MMP-3 (1:1) 37˚C, 1-2h 82 (70) 67 (64) 66.6* F88QTFE; N-term & C-term cleavage
1x Okada, JBC267/1992/21712; Shapiro JBC 270,1995,6351
MMP-10 Stromelysin-2 Like MMP-3MMP-11 Stromelysin-3 62 APMA does not
activateSantavicca Biochem J 1996 315, 953
Furin intracellularly 47 1xMMP-12 Macrophage
Metalloelastase54 Autolytic Refolding in
Ca2+/Zn2+ buffer45 22 N-tem & C- term
cleavage1x Shapiro et al., JBC 267/1992/
4664MMP-13 Collagenase-III 60 APMA, 1mM 37˚C, 30-60 min A 48 Y85NVFPRT 1x Knauper et al., JBC
271/1996/1544MMP-3 (1/10 moles) 37˚C; 3-7h CTrypsinTPCK, 10 µg/mL 37˚C: 10-30 min B
MMP-14 MT1-MMP 31rProCatDom. APMA Does not activate
25 and 23C-term trunc
Will et al., JBC 271/1996/17119
Furin IntracellularlyTrypsinTPCK, 5 µg/mL 37˚C; 10-60 min B YAIGGLKW 1x
MMP-15 MT2-MMP 33rProCatDom. Autolytic for rPro Refolding in Ca2+/Zn2+ buffer
30 24 and 22 L84 & L93 1x Kolkenbrock et al., Biol.Chem. 378/1997/ 71
MMP-16 MT3-MMP rProCatDom Autolytic for rPro Refolding in Ca2+/Zn2+ buffer
21 L91 Shofuda et al. JBC 272/1997/ 9749
MMP-17 MT4-MMP rProCatDom TrypsinTPCK, 5 µg/mL 37˚C; 30-120 min
Pei and Weiss. JBC 271/ ’96/
9135
24 Takino et al., JBC 270/1995/ 23013
MMP-24 MT5-MMP 63 nd Intracellularly 40-46 29 Pei JBC 274/1999/ 8925
activation Buffer:50mMTris;0.15MNaCl;10mMCaCl2;0.05%Brij35
activation termination: A Spin columnB PMSF2mM;Aprotinin10μg/mL;SoybeanTrypsinInhibitor2-10xoftheTrypsinamountC EDTA20mM;o-Phenantroline1mM(willinactivatetheenzymeMMPandtheactivatorMMP)
28
Cellvisualizationandmicroscopyhasremainedafundamentaltoolforinvestigatingcellularbehavior and structure, including changes in structural features as well as cellular and sub-cellular dynamics under normal and diseased states. As the technology for microscopy has evolved, the ability to interrogate discrete protein and organelle dynamics has increased and the needforspecificvisualizationtoolshasgrown.
EMD Millipore’s growing portfolio of biosensors and cell-based assays provides innovative cell visualizationandstainingsolutionsthatcomplementoursuiteoffluorescently-conjugatedprimary and secondary antibodies.
Visualization
29
FeaturedConjugatedAntibodies
anti-α-tubulin, clone dM1a, alexa Fluor® 488 conjugate(Catalogue No. 16-232)
Microtubules are constituent parts of the mitotic apparatus, cilia, flagella, and elements of
the cytoskeleton. They consist principally of 2 soluble proteins, α- and b-tubulin.
neuro-chrom™ chromaPan neuronal Marker(Catalogue No. NS420)
Neuro-Chrom™ ChromaPan Neuronal Marker is specific to axons, soma, dendrites including
spines, and nuclei of neurons.
anti-cortactin (p80/85), clone 4F11, alexa Fluor® 488 conjugate (Catalogue No. 16-228)
Cortactin, a filamentous actin-binding protein and substrate of the protein tyrosine kinase
Src, contains several potential signaling motifs. The specific tyrosine residues of Cortactin,
targeted by Src, are central to the attenuation of the F-actin cross-linking activity of this
protein.
immunocytochemistry analysis: Positive immunostaining for α-Tubulin in HeLa cells.
immunocytochemistry analysis: Positive immunostaining of rat cortex primary cells.
immunocytochemistry analysis: HeLa cells were stained with anti-Cortactin, clone 4F11, Alexa Fluor® 488 (green) and DAPI (blue).
30
lentiBrite™ lentiviral Biosensors(Multiple Catalogue Nos.)
Biosensors can be used to detect the presence/absence of a particular protein as well as the
subcellular location of that protein within the live state of a cell. Fluorescent tags are often
desired as a means to visualize the protein of interest within a cell by either fluorescent
microscopy or time-lapse video capture. Visualizing live cells without disruption allows
researchers to observe cellular conditions in real time.
Lentiviral vector systems are a popular research tool used to introduce gene products into
cells. Lentiviral transfection has advantages over non-viral methods such as chemical-based
transfection, including higher-efficiency transfection of dividing and non-dividing cells, long-
term stable expression of the transgene, and low immunogenicity.
EMD Millipore is introducing LentiBrite™ Lentiviral Biosensors, a new suite of pre-packaged
lentiviral particles encoding important and foundational proteins of autophagy, apoptosis, and
cell structure for visualization under different cell/disease states in live cell and in vitro analysis.
•Fluorescently-tagged with GFP and RFP
•Higher efficiency transfection as compared to traditional chemical-based and other
non-viral-based transfection methods
•Ability to transfect dividing, non-dividing, and difficult-to-transfect cell types such as primary
cells or stem cells
•Non-disruptive towards cellular function
Technology Highlight
LentiBrite™GFP-Tubulin(CatalogueNo.17-10206)transfection of HUVEC cells undergoing cell division.
LentiBrite™RFP-BetaActin(Catalogue No. 17-10203) transfection of HUVEC cells.
LentiBrite™GFP-Vimentin(Catalogue No. 17-10152) transfection of Human Mesenchymal Stem Cells (HuMSC).
aggregation of gFP-lc3 in autophagosomes in autophagy-induced cells. HeLa cells were transduced with lentiviral particles. Cells were either left in complete media (left) or incubated in EBSS with a lysosomal inhibitor (right) to induce autophagy and inhibit lysosomal degradation of autophagosomes. The GFP-LC3 displays a diffuse nuclear and cytosolic distribution in fed cells, and a punctate distribution in starved autophagic cells.
Watch a video of autophagy occurring in real time by scanning this QR code or by visiting: www.millipore.com/autophagyvideo
Uninduced Induced
description catalogue no.
GFP-LC3 17-10193
GFP-LC3 Mutant 17-10189
RFP-LC3 17-10143
GFP-Tubulin 17-10206
RFP-Tubulin 17-10205
GFP-Beta Actin 17-10204
RFP-Beta Actin 17-10203
GFP-Vimentin 17-10152
RFP-Vimentin 17-10153
Paxillin-GFP 17-10154
Calreticulin-RFP 17-10146
Alpha-Actinin-RFP 17-10196
lentiBrite™ lentiviral Biosensor Family
For our entire line of lentiviral biosensors, please see our website!
31
The organization of actin filaments and focal adhesions reflects cell polarity, cell cycle
state, differentiation status, and other phenotypes. This staining kit is a sensitive
immunocytochemical tool to map the local orientation of actin filaments and focal
adhesions relative to the nucleus.
actin cytoskeleton and Focal adhesion staining kit(Catalogue No. FAK100)
Confocal fluorescence microscopy of focal adhesion and actin cytoskeleton in COS-7 cells revealed with triple labeling using Phalloidin-TRITC (Orange-red), Anti-Vinculin-FITC (Green), and DAPI (Blue).
immunofluorescence analysis: Confocal fluorescence microscopy of non-treated/enriched and treated/enriched HeLa cells. (A) F-actin was detected usingTRITC-conjugated,(B)focaladhesioncontactsweredetectedusingaVinculinantibodyandaFITC-conjugatedsecondary, (C) nuclear counterstaining revealed with DAPI and all images were overlaid. Background due to soluble cytosolic fraction is significantly reduced after enrichment using the ProteoExtract® Cytoskeleton Enrichment and Staining Kit (Cat. No. 17-10210), resulting in clear detection of insoluble, low-abundance actin-associated proteins (vinculin).
Featured Product
ProteoExtract® cytoskeleton Enrichment and staining kit(Catalogue No. 17-10210)
The actin cytoskeleton serves as a scaffold to assemble multi-component signaling complexes.
Upon activation, many actin regulatory proteins/phospho-proteins move from the soluble
cytoplasmic compartment to the insoluble actin cytoskeleton. The insolubility of these
important proteins makes it difficult to study their biochemical changes upon binding to actin
including regulatory post-translational modifications (phosphorylation, nitrosylation, etc.)
EMD Millipore is introducing an easy-to-use cytoskeleton enrichment and staining kit that
quickly enriches the cytoskeleton in a gentle manner that maintains the actin cytoskeleton in
its native, adhered conformation, with minimal disruption of pre-existing protein associations.
The kit allows for the visualization and co-localization of the actin cytoskeleton and actin-
associated proteins in their native state and reduces the background typically associated with
traditional methods of whole cell staining, thus greatly increases the ability to detect and
study low abundance actin-associated proteins.
Features, Benefits, advantages:• Speeds and simplifies the enrichment of cytoskeleton-associated proteins in 20 minutes
• Maintains actin cytoskeleton and associated proteins in their native, adhered
conformations
• Significantly reduces background signal, while allowing for improved detection of low-
abundance cytoskeleton-associated proteins
description catalogue no.
ProteoExtract® Native Cytoskeleton Enrichment and Staining Kit 17-10210
ProteoExtract® Cytoskeleton Enrichment and Isolation Kit 17-10195
A
Enric
hed
cells
Non
-enr
iche
d ce
lls
usin
g 17
-102
10
B C
EMD Millipore and the M mark are trademarks and Calbiochem and ProteoExtract are registered trademarks of Merck KGaA, Darmstadt, Germany.guava, MILLIPLEX, Upstate, Chemicon, MultiScreen, and Millicell are registered trademarks of Millipore Corporation.FlowCellect, LentiBrite, QCM, AXIS, ChemiKine, ECMatrix, Milli-Mark, and Neuro-Chrom are trademarks of Millipore Corporation.Alexa Fluor is a registered trademark of Life Technologies, Inc.ReNcell is a registered trademark of ReNeuron Group plc.Luminex and xMAP are registered trademarks of Luminex Corporation.Lit No. PB1996EN00 LS-SBU-11-05319 Printed in the USA 10/2011 © 2011 Millipore Corporation. All rights reserved.
www.emdmillipore.com
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Your Source for Cell Structure ResearchWe at EMD Millipore are excited that the latest
findings in developmental biology, stem cell
research, neuroscience, and oncology have placed
cell structure and movement at the crossroads of
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For more information, please visit our website at
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