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ECM535 α/β Integrin-mediated Cell Adhesion Array Combo Kit, fluorimetric

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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ECM535
2 plates  96 wells each
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Overview

Replacement Information

Key Specifications Table

Detection Methods
Fluorescent
Description
Catalogue NumberECM535
Brand Family Chemicon®
Trade Name
  • Chemicon
Descriptionα/β Integrin-mediated Cell Adhesion Array Combo Kit, fluorimetric
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 Combo 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 Alpha/Beta Integrin-Mediated Cell Adhesion Arrays can be completed within 1-2 hours, providing a rapid, sensitive, and 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 Kit (Fluorimetric) (ECM533)
  • Beta Integrin-Mediated Cell Adhesion Array Kit (Fluorimetric) (ECM534)
  • Alpha Integrin-Mediated Cell Adhesion Array Kit (Colorimetric) (ECM530)
  • Beta Integrin-Mediated Cell Adhesion Array Kit (Colorimetric) (ECM531)
  • Alpha/Beta Integrin-Mediated Cell Adhesion Array Combo Kit (Colorimetric) (ECM532)
  • 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)
  • QCM™ 96-well Collagen-Based Cell Invasion Assay (ECM 556)
  • 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 Kit uses mouse monoclonal antibodies generated against human alpha (alpha1, alpha2, alpha3, alpha4, alpha5, alphaV, and alphavbeta3), and beta (beta1, beta2, beta3, beta4, beta6, alphaVbeta5, and alpha5beta1) 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 surface and incubated. Subsequently, unbound cells are washed away, and the adherent cells are lysed and detected by the patented CyQuant GR dye (Molecular Probes) (3-4). This green-fluorescent dye exhibits strong fluorescence enhancement when bound to cellular nucleic acids (5). Relative cell attachment is determined using a fluorescence plate reader.

    Additional information about the monoclonals provided in the kit:
    Monoclonal / Known Species Reactivity / Clone Number
    Mouse anti-Alpha1 [CD49a] / Hu / FB12
    Mouse anti-Alpha2 [CD49b] / Hu / P1E6
    Mouse anti-Alpha3 [CD49c] / Hu / P1B5
    Mouse anti-Alpha4 [CD49d] / Hu, Prim / P1H4
    Mouse anti-Alpha5 [CD49e] / Hu / P1D6
    Mouse anti-AlphaV [CD51] / Hu / P3G8
    Mouse anti-AlphaV/Beta 3 [CD51/CD61] / Hu, Bov, Por, Av, Can, Chk, Mky, Rb / LM609
    Mouse anti-Beta1 [CD29] / Hu, Mky, Rb / P4G11
    Mouse anti-Beta2 [CD18] / Hu / P4H9
    Mouse anti-Beta3 [CD61] / Hu / 25E11
    Mouse anti-Beta4 [CD104] / Hu / ASC-9
    Mouse anti-Beta6 / Hu, Rt, Mky, Rb / CSb6
    Mouse anti-AlphaV/ Beta5 [CD51/CD61] / Hu, Chk / P1F6
    Mouse anti-Alpha5/ Beta1 [VLA5, FNR] / Hu, Por / HA5
    		•
    Materials Required but Not Delivered· Multi-channel and/or repeating pipettes

    · Harvesting buffer: EDTA or trypsin cell detachment buffer. Suggested formulations include a) 2 mM EDTA/PBS, b) 0.05% trypsin in Hanks Balanced Salt Solution (HBSS) containing 25 mM HEPES, or other cell detachment formulations as optimized by individual investigators

    Note: Trypsin cell detachment buffer maybe required for difficult cell lines. Allow sufficient time for cell receptor recovery.

    · Tissue culture growth medium appropriate for subject cells, such as DMEM containing 10% FBS

    · Quenching Medium: serum-free medium, such as DMEM, EMEM, or FBM (fibroblast basal media), containing 5% BSA

    Note: Quenching Medium must contain divalent cations (Mg2+, Ca2+) sufficient for quenching EDTA in the harvesting buffer.

    · Sterile PBS or HBSS to wash cells

    · Deionized water

    · Low speed centrifuge and tubes for cell harvesting

    · CO2 incubator appropriate for subject cells

    · Sterile cell culture hood

    · Hemacytometer and microscope for counting cells

    · Trypan Blue or equivalent viability stain

    · Fluorescence plate reader with 485nm and 530nm filter set

    · 96-well plate suitable for fluorescence measurement
    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 Alpha Integrin plate layout on the insert data sheet.
    • 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 Beta Integrin plate layout on the insert data sheet.
    • 4X Cell Lysis Buffer: (Part No. 90130) One bottle - 16 mL
    • CyQuant GR Dye®: (Part No. 90132) Two vials - 75 μL each
    • Assay Buffer: (Part No. 90601) Two bottles - 100 mL each
    Detection methodFluorescent
    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 (6) 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 Alpha and Beta Integrin Array Plates can be stored at 2° to 8°C in the foil pouch up to their expiration date. 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. Keep the remaining kit components at 2° to 8°C.

    Precautions

    No data is available on the biological toxicity of CyQuant GR® dye. This reagent binds to nucleic acids, and as such, should be treated as a potential mutagen which may cause cancer and heritable genetic damage. Handle with caution. The DMSO stock solution should be handled with special caution as DMSO can facilitate the entry of organic molecules into tissues.
    Packaging Information
    Material Size2 plates
    Material Package96 wells each
    Transport Information
    Supplemental Information
    Specifications
    Global Trade Item Number
    Catalog Number GTIN
    ECM535 04053252281860

    Documentation

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

    Title

    Safety Data Sheet (SDS) 

    References

    Reference overviewPub Med ID
    Expression of matrix macromolecules and functional properties of breast cancer cells are modulated by the bisphosphonate zoledronic acid.
    P G Dedes,Ch Gialeli,A I Tsonis,I Kanakis,A D Theocharis,D Kletsas,G N Tzanakakis,N K Karamanos
    Biochimica et biophysica acta  1820  2012

    Show Abstract     		22884656 22884656

    Technical Info

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

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