The 515 monoclonal antibody specifically binds to CD44. CD44 is an 80-95 kDa type I transmembrane glycoprotein, also known as Phagocytic glycoprotein-1 (Pgp-1), or Extracellular matrix receptor type III (ECMR-III). CD44 is a member of the hyaladherin family of hyaluronan-binding proteins, with structural similarities to selectins. CD44 is the receptor for hyaluronic acid. CD44 is expressed on leucocytes, erythrocytes, epithelial cells and weakly on platelets. CD44 has functional roles in cell migration, lymphocyte homing and adhesion during hematopoiesis and lymphocyte activation. The 515 monoclonal antibody can reportedly block cellular adhesion to hyaluronic acid.
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.