Description
The 2C9.G2 monoclonal antibody specifically binds to the integrin β3 chain (CD61), which associates with the integrin αv chain (CD51) to form the vitronectin receptor, as well as the αIIb chain (CD41) to form the gpIIb/IIIa complex. Both receptors mediate adhesion to fibronectin, fibrinogen, vitronectin, thrombospondin, and von Willebrand factor. Leukocyte-endothelial adhesion is also mediated by the binding of αvβ3 integrin or vitronectin receptor to CD31 (PECAM-1). In addition, interaction of the αvβ3 integrin with its ligands regulates the L-type Ca2+ channel in vascular smooth muscle cells, possibly mediating vasodilatory responses to injury. Soluble and insoluble 2C9.G2 mAb mimics the effect of the natural ligands in smooth muscle cells from rat cremaster arterioles. Furthermore, osteopontin, also named Eta-1, is a cytokine that binds to αvβ3. CD61 is expressed on platelets, activated T lymphocytes, polymorphonuclear granulocytes, and blastocysts. Cross-reactivity of mAb 2C9.G2 to rat mast cells and platelets has been observed by flow cytometric analysis. mAb 2C9.G2 has been demonstrated to block binding of rat and mouse cells to fibronectin.
Immunohistochemistry: The 2C9.G2 antibody specific for mouse CD61 is recommended to test for immunohistochemical staining of acetone-fixed frozen sections. Tissues tested were mouse spleen and thymus. The antibody stains platelets activated T lymphocytes and polymorphonuclear granulocytes. The isotype control recommended for use with this antibody is purified hamster IgG (Cat. No. 550344). For optimal indirect immunohistochemical staining, the 2C9.G2 antibody should be titrated (1-10 to 1-50 dilution and visualized via a three-step staining procedure in combination with anti-hamster IgG cocktail (Cat. No. 550335) as the secondary antibody and Streptavidin-HRP (Cat. No. 550946) together with the DAB detection system (Cat. No. 550880). A detailed protocol of the immunohistochemical procedure is available at our website, http://www.bdbiosciences.com/support/resources
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
4. Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
5. An isotype control should be used at the same concentration as the antibody of interest.
6. This antibody has been developed for the immunohistochemistry application. However, a routine immunohistochemistry test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
