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ECM532 α/β Integrin-mediated Cell Adhesion Array Combo Kit, colorimetic

The Alpha/Beta Integrin-Mediated Cell Adhesion Array Kit can be used for assessing the presence or absence of specific integrins on the cell surface.

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ECM532
1 plate  96 wells
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      Overview

      Replacement Information

      Key Spec Table

      Detection Methods
      Chromogenic
      Description
      Catalogue NumberECM532
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      Descriptionα/β Integrin-mediated Cell Adhesion Array Combo Kit, colorimetic
      OverviewIntroduction

      Cell adhesion plays a major role in cellular communication and regulation, and is of fundamental importance in the development and maintenance of tissues. A family of cell surface receptors, the integrins, mediates cell attachment to proteins of the extracellular matrix. Integrins are heterodimer molecules containing an alpha and beta transmembrane glycoprotein subunit that are non-covalently bound together (1-2). Historically, antibodies have been used to determine the integrin profiles of cells by immunoprecipitation, immunofluorescence, immunoblotting, and flow cytometry. These methods are time-consuming, laborious or require sophisticated equipment.

      The CHEMICON® Alpha/Beta Integrin-Mediated Cell Adhesion Array kit is designed as a cost effective, efficient method for identification of cell surface integrins. The kit format uses 12 x 8-well removable strips in a plate frame for convenience and flexibility in designing assays. The Integrin-Mediated Cell Adhesion Array can be completed within 1-2 hours, providing a rapid, quantitative assay for the characterization of alpha and beta integrin surface expression.

      In addition, Chemicon continues to provide numerous migration, invasion, and adhesion products including:

      · Alpha Integrin-Mediated Cell Adhesion Array (ECM530)

      · Beta Integrin-Mediated Cell Adhesion Array (ECM531)

      · QCM™ 8μm 96-well Chemotaxis Cell Migration Assay (ECM510)

      · QCM™ 5μm 96-well Chemotaxis Cell Migration Assay (ECM512)

      · QCM™ 3μm 96-well Chemotaxis Cell Migration Assay (ECM515)

      · QCM™ 96-well Cell Invasion Assay (ECM555)

      · 24-well Insert Cell Migration and Invasion Assay Systems

      · CytoMatrix™ Cell Adhesion strips (ECM protein coated)

      · QuantiMatrix™ ECM protein ELISA kits



      Test Principle

      The CHEMICON® Alpha/Beta Integrin-Mediated Cell Adhesion Array uses mouse monoclonal antibodies generated against human alpha (alpha1, alpha2, alpha3, alpha4, alpha5, alphaV, and alphavbeta3), and beta (ßeta1, ßeta2, ßeta3, ßeta4, ßeta6, alphaVßeta5, and alpha5ßeta1) integrins/subunits, that are immobilized onto a goat anti-mouse antibody coated microtiter plate. The plate is then used to capture cells expressing these integrins on their cell surface; goat anti-mouse antibody coated wells are provided as a negative assay control. Each strip (8 wells total) consists of 7 integrin antibody coated wells and one negative control well (see plate layout on page 4). Experimental cells are seeded onto the coated substrate and incubated. Subsequently, unbound cells are washed away, and adherent cells are fixed and stained. Relative cell attachment is determined using absorbance readings.

      Additional information about the monoclonals provided in the kit:

      *Mouse anti-Alpha 1 [CD49a], clone FB12, (Hu)

      *Mouse anti-Alpha 2 [CD49b], clone P1E6, (Hu)

      *Mouse anti-Alpha 3 [CD49c], clone P1B5, (Hu)

      *Mouse anti-Alpha 4 [CD49d], clone P1H4, (Hu, Prim)

      *Mouse anti-Alpha 5 [CD49e], clone P1D6, (Hu)

      *Mouse anti-Alpha V [CD51], clone P3G8, (Hu)

      *Mouse anti-Alpha V/Beta 3 [CD51/CD61], clone LM609, (Hu, Bov, Por, Av, Can, Chk, Mky, Rb)

      *Mouse anti-Beta 1 [CD29], clone P4G11, (Hu, Mky, Rb)

      *Mouse anti-Beta 2 [CD18], clone P4H9, (Hu)

      *Mouse anti-Beta 3 [CD61], clone 25E11, (Hu)

      *Mouse anti-Beta 4 [CD104], clone ASC-9, (Hu)

      *Mouse anti-Beta 6, clone CSb6, (Hu, Rt, Mky, Rb)

      *Mouse anti-Alpha V/ Beta 5 [CD51/CD61], clone P1F6, (Hu, Chk)

      *Mouse anti-Alpha 5/ Beta 1 [VLA5, FNR], clone HA5, ( Hu, Por)
      Materials Required but Not Delivered· Multi-channel or repeating pipettes

      · Microplate reader (540-570 nm detection) or spectrophotometer

      · CO2 incubator appropriate for subject cells

      · Hemacytometer and microscope for counting cells

      · Dissociation Buffer

      · Deionized water

      · PBS
      References
      Product Information
      Components
      • Alpha Integrin Array Plate: (PN: 90599) One 96-well plate with 12 strips. Each strip contains one well each of the mouse anti-alpha integrin monoclonal antibodies: (alpha1, alpha2, alpha3, alpha4, alpha5, alphaV, alphavbeta3), and one goat anti-mouse negative well. See the plate layout on product insert.
      • Beta Integrin Array Plate: (PN: 90600) One 96-well plate with 12 strips. Each strip contains one well each of the mouse anti-beta integrin monoclonal antibodies: (beta1, beta2, beta3, beta4, beta6, alphaVbeta5, and alpha5beta1), and one goat anti-mouse negative well. See the plate layout on product insert.
      • Cell Stain Solution: (Part No. 90144) Two 20 mL bottles.
      • Extraction Buffer: (Part No. 90145) Two 20 mL bottles.
      • Assay Buffer: (Part No. 90601) Two 100 mL bottles.
      Detection methodChromogenic
      Quality LevelMQ100
      Applications
      ApplicationThe Alpha/Beta Integrin-Mediated Cell Adhesion Array Kit can be used for assessing the presence or absence of specific integrins on the cell surface.
      Application NotesThe CHEMICON® Alpha/Beta Integrin-Mediated Cell Adhesion Array Kit can be used for assessing the presence or absence of specific integrins on the cell surface. This assay can substitute for FACS analysis (3) and is useful for screening human adherent cells for cell surface integrins or for monitoring in vitro cell differentiation or genetic modification of cells.



      For Research Use Only. Not for use in diagnostic procedures.
      Biological Information
      Species Reactivity
      • Human
      Entrez Gene Number
      Entrez Gene SummaryThis gene encodes the alpha 1 subunit of integrin receptors. This protein heterodimerizes with the beta 1 subunit to form a cell-surface receptor for collagen and laminin. The heterodimeric receptor is involved in cell-cell adhesion and may play a role in inflammation and fibrosis. The alpha 1 subunit contains an inserted (I) von Willebrand factor type I domain which is thought to be involved in collagen binding.
      Gene Symbol
      • ITGA1
      • CD49a
      • VLA1
      • VLA-1
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P56199 # Integrin alpha-1/beta-1 is a receptor for laminin and collagen. It recognizes the proline-hydroxylated sequence G-F-P-G- E-R in collagen.
      SIZE: 1179 amino acids; 130848 Da
      SUBUNIT: Heterodimer of an alpha and a beta subunit. Alpha-1 associates with beta-1.
      SUBCELLULAR LOCATION: Membrane; Single-pass type I membrane protein.
      DOMAIN: SwissProt: P56199 The integrin I-domain (insert) is a VWFA domain. Integrins with I-domains do not undergo protease cleavage.
      SIMILARITY: Belongs to the integrin alpha chain family. & Contains 7 FG-GAP repeats. & Contains 1 VWFA domain.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsThe experimental and control plates can be stored at 2° to 8°C in the foil pouch up to their expiration. Unused strips may be placed back in the pouch for storage. Ensure that the desiccant remains in the pouch, and that the pouch is securely closed.

      Precautions

      · Cell stain contains minor amounts of crystal violet, a toxic substance, which may cause cancer and heritable genetic damage. Handle with caution. Toxic by inhalation and if swallowed. Irritating to eyes, respiratory system and skin.

      · Extraction buffer contains alcohol. Avoid internal consumption.
      Packaging Information
      Material Size1 plate
      Material Package96 wells
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      ECM532 04053252323928

      Documentation

      α/β Integrin-mediated Cell Adhesion Array Combo Kit, colorimetic SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewPub Med ID
      Effects of amelogenins on angiogenesis-associated processes of endothelial cells.
      S Almqvist,H K Kleinman,M Werthén,P Thomsen,M S Agren
      Journal of wound care  20  2011

      Show Abstract       		21378680 21378680
      Activin A regulates trophoblast cell adhesive properties: implications for implantation failure in women with endometriosis-associated infertility.
      Chelsea J Stoikos,Lois A Salamonsen,Natalie J Hannan,Anne E O'Connor,Luk Rombauts,Evdokia Dimitriadis
      Human reproduction (Oxford, England)  25  2010

      Show Abstract       		20457668 20457668

      Technical Info

      Title
      Integrin Profiling of Stem Cells and Differentiated Progeny
      Screening Kits to Monitor Cell - Extracellular Matrix Interactions

      FAQ

      QuestionAnswer
      When you say wash gently, what do you mean? How is this different from normal washing?To wash gently, this is what we would recommend: remove all of the initial solution but 50 microliters. Then, add 250 microliters of wash. With your pipette set at 150 microliters, pipette up and down. Move the plate around and wash all sides of the well to ensure uniformity. Remove the 150 microliters, then add another 150 microliters, repeating this wash step five times.

      Between washes, you may want to check the test and control wells under the microscope to ensure that you are not washing too vigorously or too gently. When the control wells have about 10% or less cells still attached, read your plate. Of course, washing stringency will depend on the cell type as they differ in their binding capacities, and this should be determined by the researcher.

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      Categories

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