The BMA-12 monoclonal antibody specifically binds to CD100 which is also known as, Semaphorin-4D (Sema4D), or Collapsin-4 (Coll-4). CD100 is a ~150 kDa type I transmembrane glycoprotein that belongs to the class IV Semaphorin subfamily within the Ig gene superfamily. CD100 is expressed by cells from a broad range of tissues including the nervous system, where it serves in axon guidance and synapse formation, and the kidney. It is expressed by most hematopoietic cells including T cells which express CD100 more abundantly than B cells and dendritic cell subsets. Its expression is significantly upregulated upon activation of these cell types. CD100 can costimulate T cell responses. For B cells and dendritic cells, CD100 interaction with its ligands, CD72 or Plexin B1, induces cellular activation and promotes survival, respectively. CD100 is also known to interact with Plexin B2 on skin gamma delta T cells and contribute to the repair of tissue damage caused by inflammation. A biologically-active, soluble form of CD100 is generated by proteolytic cleavage of the cell surface form.
The antibody was conjugated to BD Horizon™ BUV563 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 which has an Ex Max of 348 nm and an acceptor dye. The tandem has an Em Max at 563 nm. BD Horizon BUV563 can be excited by the 355 nm ultraviolet laser. On instruments with a 561 nm Yellow-Green laser, the recommended bandpass filter is 585/15 nm with a 535 nm long pass to minimize laser light leakage. When BD Horizon BUV563 is used with an instrument that does not have a 561 nm laser, a 560/40 nm filter with a 535 nm long pass may be more optimal. Due to the excitation and emission characteristics of the acceptor dye, there may be spillover into the PE and PE-CF594 detectors. However, the spillover can be corrected through compensation as with any other dye combination.