The 34-2-12 antibody (also known as 34-2-12S) recognizes the α3 domain of the H-2D[d]. The binding of the antibody to its epitope is independent of the α1 and α2 domains and β2 microglobulin. It cross-reacts with cells of the C3H.LG/Ckc strain. Reactivity with other haplotypes (eg, b, f, k, p, q, r, s) has not been observed. Soluble mAb 34-2-12 blocks binding of the Ly-49A-expressing T lymphoma EL4 to immobilized H-2D[d]. However, further studies utilizing this mAb indicate that the α3 domain is not involved in the interaction between Ly-49A, or Ly-49G2, and H-2D[d].
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.