Description
The MBC 78.2 monoclonal antibody recognizes CD31 which is also known as, Platelet endothelial cell adhesion molecule (PECAM-1), platelet GPIIa, or EndoCAM. CD31 is a ~130 kDa type I transmembrane glycoprotein that belongs to the Ig gene superfamily. CD31 is comprised of an extracellular region with six IgC-like domains, a transmembrane region, and a cytoplasmic domain that contains two immunoreceptor tyrosine-based inhibitory motifs (ITIMs). The MBC 78.2 antibody specifically binds to an epitope located on membrane-proximal, extracellular Ig-like domain 6 of CD31. This epitope remains expressed by activated T cells after enzymatic cleavage and shedding of a soluble extracellular CD31 fragment comprised of Ig-like domains 1 to 5 from cells. In contrast to the MBC 78.2 antibody, the WM59 monoclonal antibody reportedly binds to the extracellular Ig-like domain 2 of CD31. WM59 can thus bind to cells that express intact CD31 but not to cells that express a truncated form CD31 that lacks at least the membrane distal Ig-like domains 1 and 2 of CD31. CD31 has wide tissue distribution and is expressed on platelets, monocytes, granulocytes, some T cell subsets, and at high levels on endothelial cells. This cell adhesion molecule has been implicated in a number of cellular phenomena, including vascular wound healing. angiogenesis, transendothelial migration of leucocytes, and the regulation of T cell responses.
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. 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.
1. 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.
2. Researchers should determine the optimal concentration of this reagent for their individual applications.
3. 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.
4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
5. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
6. An isotype control should be used at the same concentration as the antibody of interest.
7. 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.
8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
10. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
11. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
12. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
