The AD2 monoclonal antibody specifically binds to ecto-5'-nucleotidase, a 70 kDa, glycosyl phosphatidylinositol (GPI)-anchored glycoprotein. CD73 is expressed on subsets of T and B lymphocytes, follicular dendritic cells, epithelial cells, endothelial cells and mesenchymal stem cells. Its expression on lymphocytes increases during T and B cell development. CD73 has enzymatic activity and catalyzes the dephosphorylation of adenosine monophosphate (AMP) converting it to adenosine. It has been suggested that CD73 can mediate costimulatory signals in T cell activation and adhesion of lymphocytes to endothelium.
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.