Anti-Integrin alphaV Antibody, CT, Intracellular

The integrin family of cell adhesion receptors consists of at least 19 membrane-associated heterodimers, composed of an alpha and beta subunit that associate in a non-covalent manner. To date, 16 different alpha-subunits and 8 different beta-subunits have been identified. The structure and functional diversity of the integrin family are based upon the pairing abilities of the individual alpha and beta subunits. Key to these molecular interactions between the integrin receptors and their respective ligands is the recognition of the Arg-Gly-Asp (RGD) sequence, known to be present in the extracellular matrix components fibronectin, vitronectin, collagen, fibrinogen, and von Willebrand factor (Cheresh, 1991). The involvement of integrins in vascular proliferation, adhesion, and wound repair has been well documented. Integrin alphaV is a type I membrane glycoptroein with no I (inserted) domain. The alphaV protein is post-translationally cleaved into a heavy and light chain, which is expressed on the cell surface as a disulfide linked polypeptide.

Integrin alphaV. Specificity determined by immunoprecipitation from a detergent extract of 35S methonine labeled endothelial cells. No cross-reactivity to alpha1, alpha2, alpha3, alpha4, or alpha6 integrin subunits was detected. The mature alphaV protein (150 kDa) is composed of a disulfide bonded heavy (125 kDa) and a light (25 kDa) chain derived from a proteolytically cleaved precursor (Suzuki, 1987). The epitope recognized by the antibody is located on the COOH terminal portion of the light chain. By western blot under reducing conditions the antibody detects two major bands at approximately 25 and 27 kDa corresponding to the light chain. The slight differences in sizes of the light chains is thought to be caused by proteolytic cleavage at two alternative sites on the precursor. Similar sites have been reported for alpha6, alpha3, and alphalIbeta (Hogervorst, 1991).

Epitope: C-terminus, intracellular

Synthetic peptide derived from the COOH terminal sequence (cytoplasmic domain) of the human alphaV integrin subunit (SwissProt Accession P06756, amino acids 1022-1034).

Anti-Integrin alphaV Antibody, C-terminus, Intracellular detects level of Integrin alphaV & has been published & validated for use in ELISA, IH, IP, RIA & WB.

ELISA/RIA:
A 1:1,000 dilution of a previous lot was used in ELISA/RIA.

Immunoprecipitation:
5 μL of a previous lot of this antibody is sufficient to precipitate alphaV/beta1 from 5x106 cells.

Immunohistochemistry:
1:1000 dilution from a previous lot was used in immunohistochemistry. (Suggested for use an acetone fixed tissue only.)

Western blot: 1:5000.
The antibody recognizes a single band at approximately 150 kDa when denatured, but not reduced (Hirsch, 1994). Under reducing conditions this antibody recognizes two major bands of 25 and 27 kDa and, to a lesser extent, the full length 150 kDa band.

Optimal working dilutions must be determined by end user.

Research Category
Cell Structure

Research Sub Category
Integrins

Routinely evaluated by Western Blot on C6 lysates.
Western Blot Analysis: 1:500 dilution of this lot detected INTEGRIN AV light chain on 10 μg of C6 lysates.

130 kDa; 25 and 27 kDa (reducing conditions)

Neat rabbit antiserum in liquid with 0.05% sodium azide.

Unpurified

Stable for 1 year at -20°C in undiluted aliquots from date of receipt.

Handling Recommendations:
Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Control
HT-29 colon carcinoma cells

C6 lysates.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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