The 1-1-2 monoclonal antibody specifically recognizes the Qa-2 MHC class Ib antigen, which is expressed at higher levels on mature T lymphocytes than on B cells. Its expression is upregulated in the late stages of T-cell maturation. Qa-2 is a 40 kDa GPI-linked cell surface molecule encoded by the Q5, Q6, Q7, and/or Q9 genes of the H-2 complex. A 39 kDa soluble form results from alternative splicing of the transcript. Although Qa-2 is nonpolymorphic, its level of expression on both T and B lymphocytes varies among inbred mouse strains. High expressors include strains carrying the H-2[b] haplotype low expressors include H-2[d] strains, and H-2[k] strains are Qa-2-negative.
The antibody was conjugated to BD Horizon™ BUV661 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 661-nm. BD Horizon Brilliant BUV661 can be excited by the ultraviolet laser (355 nm) and detected with a 670/25 filter and a 630 nm LP. Due to cross laser excitation of this dye, there may be significant spillover into channels detecting APC-like emissions (eg, 670/25-nm filter).
Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV661 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV661 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone-specific compensation controls when using these reagents.