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

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

Chemicon (Millipore)

Introduction

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)

The 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.

Human

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.

Cell Based Assays

• 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.

The 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.

Chromogenic

96 wells

• ITGA1
• CD49a
• VLA1
• VLA-1

Integrins, ECM, & Adhesion

· 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