The 2D3 monoclonal antibody specifically binds to CD249. CD249 is encoded by ENPEP and is also known as Aminopeptidase A (APA/AP-A), Glutamyl aminopeptidase (EAP), and gp160. CD249 is expressed by epithelial and endothelial cells in a variety of tissues including those that comprise the proximal tubules and glomeruli of the kidneys. It is also expressed by early B-lineage cells and cell lines. CD249 belongs to the peptidase M1 family and catalyzes the release of N-terminal glutamate (and to a lesser degree, aspartate) from peptides. This ectoenzyme catalyzes the conversion of angiotensin II to angiotensin III which is important in the local regulation of blood pressure. It may also play roles in the regulation of angiogenesis and development of B cells.
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.