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ECM531 β Integrin-mediated Cell Adhesion Array Kit, colorimetric

This assay can substitute for FACS analysis & is useful for screening human adherent cells for cell surface integrins or for monitoring in vitro cell differentiation or genetic modification of cel

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ECM531
1 plate  96 wells
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Overview

Key Spec Table

Detection Methods
Chromogenic
Description
Catalogue NumberECM531
Brand Family Chemicon®
Trade Name
  • Chemicon
Descriptionβ Integrin-mediated Cell Adhesion Array Kit, colorimetric
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® 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 beta integrin surface expression.

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

· Alpha Integrin-Mediated Cell Adhesion Array (ECM530)

· Alpha/Beta Integrin-Mediated Cell Adhesion Array Combo Kit (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)

· 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® Beta Integrin-Mediated Cell Adhesion Array uses mouse monoclonal antibodies generated against human beta integrins/subunits (Ãeta1, Ãeta2, Ãeta3, Ãeta4, Ãeta6, alphaVÃeta5, and alpha5Ãeta1), 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 3). 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-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
Product Information
Components
  • 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) One 20 mL bottle.
  • Extraction Buffer: (Part No. 90145) One 20 mL bottle.
  • Assay Buffer: (Part No. 90601) One 100 mL bottle.
Detection methodChromogenic
Quality LevelMQ100
Applications
ApplicationThis assay can substitute for FACS analysis & is useful for screening human adherent cells for cell surface integrins or for monitoring in vitro cell differentiation or genetic modification of cells.
Application NotesThe CHEMICON® 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 SummaryIntegrins are heterodimers comprised of alpha and beta subunits, that are noncovalently associated transmembrane glycoprotein receptors. Different combinations of alpha and beta polypeptides form complexes that vary in their ligand-binding specificities. Integrins mediate cell-matrix or cell-cell adhesion, and transduced signals that regulate gene expression and cell growth. This gene encodes the integrin beta 4 subunit, a receptor for the laminins. This subunit tends to associate with alpha 6 subunit and is likely to play a pivotal role in the biology of invasive carcinoma. Mutations in this gene are associated with epidermolysis bullosa with pyloric atresia. Multiple alternatively spliced transcript variants encoding distinct isoforms have been found for this gene.
Gene Symbol
  • ITGB1
  • GPIIA
  • VLAB
  • OTTHUMP00000063732
  • MDF2
  • FNRB
  • OTTHUMP00000063731
  • OTTHUMP00000046253
  • CD29
  • OTTHUMP00000063733
  • MSK12
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P18564 # Integrin alpha-V/beta-6 is a receptor for fibronectin and cytotactin. It recognizes the sequence R-G-D in its ligands.
SIZE: 788 amino acids; 85936 Da
SUBUNIT: Heterodimer of an alpha and a beta subunit. Beta-6 associates with alpha-V. Interacts with FLNB.
SUBCELLULAR LOCATION: Membrane; Single-pass type I membrane protein.
SIMILARITY: SwissProt: P18564 ## Belongs to the integrin beta chain family. & Contains 1 VWFA domain.
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
Global Trade Item Number
Catalogue Number GTIN
ECM531 04053252734724