Ly-49C and Ly49I Mouse anti-Mouse, Brilliant Violet 650, Clone: 5E6., BD Optibuild™

Description

• The 5E6 (also known as clone SW5E6) antibody reacts with Ly-49C[BALB], Ly-49C[B6], Ly-49C[NZB], and Ly-49I[B6], inhibitory receptors which are expressed on subsets of natural killer (NK) cells and NK-1.1+ (or DX5+) T lymphocytes (NK-T cells) in all strains tested except C57BR and RIII, on a population of memory CD8+ T lymphocytes and NK1.1+ T cells in C57BL/6 mice, and on a distinct subset of B-1 cells of BALB/c and C57BL/6 mice
• The proportion of NK T cells expressing Ly-49C/I is higher (2-5 fold) in thymus than in liver (immature and mature NK T cells, respectively), and there is evidence that the down-regulation of Ly-49 receptor expression is necessary for normal NK T-cell development
• Most NK cells express a single allele of Ly-49C, although occasionally they may express more than one allele
• The Ly-49 family of NK-cell receptors are disulfide-linked type-II transmembrane protein homodimers with extracellular carbohydrate-recognition domains (CRD) that bind to MHC class I alloantigens
• 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 5E6 antibody is specific for the Ly-49C CRD
• The Ly-49C[BALB] and Ly-49C[B6] alloantigens bind to MHC class I antigens of the b, d, k, and s haplotypes, and the 5E6 antibody blocks this binding
• Binding of Ly-49C[BALB]- and Ly-49C[B6]- expressing transfectants to lymphoblasts of H-2[f], H-2[q], H-2[r], and H-2[v] strains has also been detected
• Ly-49I[B6] transfectants bind H-2[r] lymphoblasts and bind much more weakly to the b, d, k, q, s, and v haplotypes
• The levels of the Ly-49 inhibitory receptors are down-regulated by their ligands in vivo, and the various levels of expression of an Ly-49 inhibitory receptor may affect the specificity of NK cells
• Ly-49C is specifically downregulated in the presence of H-2K[b] class I molecules (one of the Ly-49C ligands)
• Ly-49C[+] and/or Ly-49I[+] cells mediate allogeneic and hybrid resistance to H-2d bone marrow transplantation
• The antibody was conjugated to BD Horizon™ BV650 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 405nm and an acceptor dye with an Em Max at 650nm
• BD Horizon BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20nm filter)
• Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor™ 700 detectors
• However, the spillover can be corrected through compensation as with any other dye combination
• BV650 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an emission maximum at 650 nm
• Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor™ 700 detectors
• BV650 will have moderate spillover into the BD Horizon™ BV711 detector.