The 14-4-4S monoclonal antibody specifically recognizes the I-E[k] MHC class II alloantigen. It crossreacts with cells from mice of the H-2[d], H-2[p], and H-2[r] haplotypes. Cells from mice of the H-2[b], H-2[f], H-2[g7], H-2[q], and H-2[s] haplotypes do not express I-E antigen. It has been reported that mAb 14-4-4S crossreacts with the rat MHC class II antigen RT1D.
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.