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

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-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

to Place an order or receivetechnical assistanceIn the U.S. and Canada, call toll-free 1-800-645-5476

For other countries across Europe and the world, please visit www.millipore.com/offices

For Technical Service, please visit www.millipore.com/techservice

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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

so many diverse research areas. As the only life

science supplier to be a steady partner to all these

fields, we are committed to providing the most

complete solutions for advancing your research.

For more information, please visit our website at

www.millipore.com/cellstructure.