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™ BUV496 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 496-nm. BD Horizon BUV496 can be excited by the ultraviolet laser (355 nm) and detected with a 515/30 nm filter with a 450LP. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into the channel detecting BD Horizon V500 or BV510 (eg, 525/40-nm filter). However, the spillover can be corrected through compensation as with any other dye combination.