The 28-14-8 antibody reacts with the α3 domain of the H-2D[b] MHC class I alloantigen. The antibody binds to H-2D[b] in the presence or absence of the β2 microglobulin chain. It cross-reacts with the α3 domain of H-2L[d], but not with K[d] or D[d], and with H-2D[q] and/or L[q]. Reactivity with haplotypes k, f, p, r, and s has not been observed. mAb 28-14-8 has been shown to block H-2L[d]- specific and H- 2L[d]-restricted antigen-specific lysis of target cells by cytotoxic T lymphocytes (CTL), but it does not block recognition of H-2L[d] positive target cells by Ly-6G2-positive NK cells.
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.