The 20d5 monoclonal antibody specifically recognizes NKG2A, NKG2C, and NKG2E (also known as CD159a, CD159c, and CD159e which are encoded by Klrc1, Klrc2, and Klrc3, respectively) on a subset of NK and NK-T cells in most strains tested (eg, AKR/J, BALB/c, C3H/He, C57BL/6, CBA/J, DBA/1, FVB/N, 129/Sv, NOD, SWR, and most DBA/2 substrains, but not DBA/2J). The NKG2 molecules are a family of lectin-like receptors that form heterodimers with CD94 on the surface of NK cells. DBA/2J mice do not express CD94, and the lack of CD94 is responsible for the absence of NKG2 expression in this substrain. NKG2 receptors are also expressed on CD8+ T lymphocytes activated in vivo and in vitro. The heterodimers of CD94 with NKG2A, C, or E recognize Qa-1, a nonclassical MHC class I antigen, presenting the Qdm peptide. Studies of CD94/NKG2 heterodimers on human NK cells have demonstrated that the NKG2 components mediate signal transduction for the receptor, with NKG2A being inhibitory and NKG2C being stimulatory. The CD94/NKG2E heterodimer is also thought to be stimulatory. The mouse NKG2A molecule contains two intracytoplasmic sequences that resemble the ITIM (Immunoreceptor Tyrosine- based Inhibitory Motif) consensus sequence. NKG2A transcripts have been shown to be up to 20-fold more abundant than NKG2C and NKG2E mRNA in NK cells of adult mice. The CD94/NKG2 receptors show increased expression on neonatal NK cells compared to the Ly-49 MHC class I receptors, suggesting that CD94/NKG2 receptors and their ligand, Qa-1, may play a role in maintenance of self-tolerance in developing NK cells. The 20d5 antibody is useful for identification of NK cells expressing functional CD94/NKG2 receptors, in contrast to the non-functional CD94 expressed alone, and it blocks the binding of Qdm-complexed Qa-1b tetramers to CD94/NKG2-transfected CHO cells.
The antibody was conjugated to BD Horizon™ BV711 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 711-nm. BD Horizon BV711 can be excited by the violet laser and detected in a filter used to detect Cy™5.5 / Alexa Fluor® 700-like dyes (eg, 712/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there may be moderate spillover into the Alexa Fluor® 700 and PerCP-Cy5.5 detectors. However, the spillover can be corrected through compensation as with any other dye combination.