The HBF-719 antibody reacts with the B6 alloantigen of Ly-49F, an inhibitory receptor which is expressed on subsets of natural killer (NK) cells, NK-1.1+ (or DX5+) T lymphocytes (NK-T cells), and on a population of memory CD8+ T lymphocytes in C57BL/6 mice, 1 but not AKR/J, BALB/c, C3H/HeN, DBA/1, or SJL mice.3 The Ly-49 family of NK-cell receptors are disulfide linked type-II transmembrane protein homodimers with extracellular carbohydrate recognition domains (CRD). The Ly-49 family members are expressed independently, such that an individual NKor T-cell may display more than one class of Ly-49 receptor homodimers. Ly-49F has an ITIM (Immunoreceptor Tyrosine-based Inhibitory Motif) in its cytoplasmic tail. Therefore, it is probably an inhibitory receptor. It weakly binds to cells expressing the H-2d MHC class I alloantigens.
The antibody was conjugated to BD Horizon BUV615 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 615 nm. BD Horizon Brilliant BUV615 can be excited by the ultraviolet laser (355 nm) and detected with a 610/20 filter and a 595 nm LP. Due to the excitation of the acceptor dye by the blue/yellow-green laser line, there may be significant spillover into channels detecting PE-CF594 like emissions (eg, 610/20-nm filter).