The G28-8 monoclonal antibody specifically recognizes RP105/Bgp95, a 95-105 kDa type I membrane protein consisting of extracellular leucine-rich repeats and a short cytoplasmic domain. It is expressed on mantle zone B cells, but weakly on germinal center B cells. RP105/Bgp95 is also expressed on peripheral blood monocytes, dendritic cells, and a subset of peripheral blood lymphocytes. The extracellular domain associates with a molecule called MD-1 to form a cell surface receptor complex RP105/Bgp95/MD- 1. This receptor belongs to the family of toll-like receptors (TLR). Studies show that RP105/Bgp95/MD-1, working in concert with TLR4, controls B cell recognition and signaling of lipopolysaccharide (LPS). Reports on functional studies show that G28-8 monoclonal antibody can induce a G0 to G1 cell cycle transition and was synergistic with PMA, anti-µ, or anti-CD40 in inducing proliferation of resting B cells.
The antibody was conjugated to BD Horizon™ BV650 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm. BD Horizon BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor® 700 detectors. However, the spillover can be corrected through compensation as with any other dye combination.