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.
Bead-based compensation or unmixing controls, such as BD® CompBeads or BD™ SpectraComp™, can be used as surrogates to assess fluorescence spillover when bound to fluorochrome-conjugated antibodies. Although these beads have spectral properties similar to cells, variations in spectral emission may occur, resulting in differing spillover values compared to biological controls. Therefore, it is considered best practice to compare the spillover obtained from cells and bead-based controls when using BD® CompBeads or BD™ SpectraComp™ for the first time, to ensure they are appropriate for the intended application.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
3. For U.S. patents that may apply, see bd.com/patents.
4. 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.
5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
6. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
7. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
9. An isotype control should be used at the same concentration as the antibody of interest.
10. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
11. Alexa Fluor™ is a trademark of Life Technologies Corporation.
12. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
