The IK22-5 monoclonal antibody specifically recognizes CD254, which is also known as, Receptor Activator of NF-kB Ligand (RANKL), TNF-Related Activation-Induced Cytokine Receptor (Trance), Osteoprotegerin Ligand (OPGL), or Osteoclast Differentiation Factor (ODF). CD254/RANKL is likewise known as Tnfsf11, a member of the tumor necrosis factor family. RANKL has been implicated in interactions between T cells and dendritic cells. RANKL binds to RANK expressed on dendritic cells and upregulates the expression of anti-apoptotic protein BcL-XL suggesting a role in dendritic cell survival. RANKL is also important in T- and B-cell maturation. RANKL can both activate mature osteoclasts and mediate osteoclastogenesis. OPGL-deficient mice show severe osteoporosis and complete absence of osteoclasts as a result of lack of osteogenesis. It is expressed abundantly by mouse activated T cells, not B cells. It is also expressed on osteoclasts.
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.