The HI30 monoclonal antibody specifically binds to the 180, 190, 205, 220 kDa protein isoforms of CD45. CD45 is encoded by the PTPRC (Protein tyrosine phosphatase receptor type C) gene. CD45, also known as the leukocyte common antigen (LCA), is a member of the protein tyrosine phosphatase (PTP) family. It is present on all human leukocytes including lymphocytes, monocytes, granulocytes, eosinophils, and thymocytes. CD45 is absent from circulating erythrocytes, platelets, or mature erythroid cells of bone marrow and non-hemopoietic tissues.
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.