Clone M-T477 reacts with the human form of a 56 kDa transmembrane glycoprotein, CD4, present on the T-helper/inducer subset of normal human donor peripheral blood lymphocytes. This clone also cross-reacts with a subset of CD3-positive peripheral blood lymphoyctes, but not monocytes, of both rhesus and cynomolgus macaque monkeys. Cross-reactivity on both lymphocytes and monocytes (weak) of baboon is also observed. The distribution of M-T477 reactivity on lymphocytes is similar for both human and monkey, with the majority of CD4-positive lymphocytes being CD8-negative and lacking reactivity with antibodies to B- or NK-cell markers.
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.