CD21 is a 145 kDa transmembrane glycoprotein that is expressed on mature B cells, follicular dendritic cells, a subset of immature thymocytes and on some epithelial cells. CD21 is also known as the C3d Receptor, Complement Receptor 2 (CR2), and the Epstein-Barr Virus receptor, (EBV-R). CD21 is part of a large signal-transduction complex that also includes CD19, CD81 and Leu13. CR2 serves as a receptor for the activation fragments of complement component C3, i.e. C3d, iC3d, and C3dg and C3d-bound antigens and plays a central role in the immune response by amplifying B-cell activation and proliferation. CR2 is also the receptor for EBV by binding its surface glycoprotein gp350/220. Anti-human CR2 hybridomas were generated from Cr2-/-mice (lacking the cr2 gene, that encodes CR2 and CR1 in mice) immunized with the recombinant protein representing the first two amino-terminal short consensus repeats (SRC) of human CR2, SCR1-2. Both the C3d and EBV binding sites of hu CR2 were localized to these sites. Clone 1048 has been reported to specifically react with CR2-expressing cells and to inhibit CR2 interactions with C3 fragments or the EBV surface protein, gp350/220.
The antibody was conjugated to BD Horizon™ BUV661 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 661-nm. BD Horizon Brilliant BUV661 can be excited by the ultraviolet laser (355 nm) and detected with a 670/25 filter and a 630 nm LP. Due to cross laser excitation of this dye, there may be significant spillover into channels detecting APC-like emissions (eg, 670/25-nm filter).
Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV661 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV661 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone-specific compensation controls when using these reagents.