The A1 monoclonal antibody specifically binds to the Ly-49A[B6] alloantigen, an inhibitory receptor that is expressed on subsets of natural killer (NK) cells and NK-1.1-positive T lymphocytes (NKT cells) in C57BL/6, C57BL/10, and B10 congenic mice, on a population of memory CD8+ T lymphocytes and NK1.1+ γδ T cells in C57BL/6 mice, and on a distinct subset of B-1 cells (CD5+B220[lo]) of C57BL/6 mice. The A1 antibody has also been reported to crossreact with Ly-49ANOD, Ly-49PNOD, Ly-49P129/J, and Ly-49V129/J alloantigens. The proportion of NKT cells expressing Ly-49A is higher (2-5 fold) in thymus than in liver (immature and mature NKT cells, respectively), and there is evidence that the down regulation of Ly-49 receptor expression is necessary for normal NKT cell development to occur. Most NK cells express a single allele of Ly-49A, although occasionally they may express more than one allele. The Ly-49 family of NK-cell receptors, members of the C-type lectin superfamily, are disulfide-linked type-II transmembrane protein homodimers with extracellular carbohydrate-recognition domains (CRD) that bind to MHC class I alloantigens. The A1 antibody is specific for the Ly-49A[B6] CRD. The Ly-49 family members are expressed independently, such that an individual NK or T cell may display more than one class of Ly-49 receptor homodimers. The Ly-49A[B6] allonantigen binds to H-2D[d], H-2D[k], and H-2D[p], and the A1 antibody blocks this binding. Binding of Ly-49A[B6] to lyphoblasts expressing MHC class I antigens of the f, q, r, s, and v haplotypes has also been demonstrated. The levels of the Ly-49 inhibitory receptors are down-regulated by their ligands in vivo, and various levels of expression of a Ly-49 inhibitory receptor may affect the specificity of NK cells. In vitro studies suggest that the Ly-49A receptor mediates negative regulation of NK-cell cytolytic activity via tyrosine phosphorylation of its ITIM (Immunoreceptor Tyrosine-based Inhibitory Motif).
The antibody was conjugated to BD Horizon™ BUV395 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye has been exclusively developed by BD Biosciences to have minimal spillover into other detectors, making it an optimal choice for multicolor flow cytometry. With an Ex Max at 348 nm and an Em Max at 395 nm, BD Horizon BUV395 can be excited with a 355 nm laser and detected with a 379/28 filter.