The VIM3b monoclonal antibody recognizes CD97. CD97 is a seven-span transmembrane protein that belongs to the G-protein coupled receptor superfamily. CD97 is encoded by ADGRE5 (adhesion G protein-coupled receptor E5). CD97 is highly expressed on monocytes and granulocytes. It is also expressed on dendritic cells, T cells, B cells, NK cells, and smooth muscle cells. CD97 expression is upregulated on activated T cells. CD97 is also expressed on several cell lines including HL-60, THP-1, and K562. CD97 reportedly plays roles in leucocyte signaling, adhesion, and migration. The VIM3b antibody works in Western blotting and immunoprecipitation. The antibody recognizes proteins of 74, 80 and 89 kDa. CD55 and chondroitin sulfate are ligands for CD97.
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.