Ly-49D Rat anti-Mouse, Brilliant Violet 650, Clone: 4E5., BD Optibuild™

Description

• The 4E5 monoclonal antibody specifically recognizes Ly-49D, which is expressed on subsets of natural killer (NK) cells in C57BL/6, C3H/He (at a very low frequency), and SJL, but not DBA/2, AKR, CBA/J, or BALB/c mice
• Unlike Ly-49D antigen has not been betected on NK-1.1+ (or DX5+) T cells
• In 129/J mice, the 4E5 antibody cross-reacts with Ly-49O, Ly-49R, and Ly-49V
• 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, which 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
• Ly-49D weakly binds to MHC class I antigens of the k halpotype, and Ly-49D+ IL-2-activated NK cells lyse target cells expressing H-2[a], H-2[b], H-2[d], H-2[k], H-2[p], H-2[q], and H-2[s] and the CHO (Chinese hamster ovary) cell line
• Ly-49D+ cells mediate allogenic resistance to H-2d bone marrow transplantation
• In vitrostudies suggest that the Ly-49D receptor mediates activation of NK-cell cytolytic activity via tyrosine phosphorylation of their ITIMs ( Immunoreceptor Tyrosine-based Inhibitory Motifs)
• Molecular differences between the Ly-49D stimulatory receptor and the inhibitory members of the Ly-49 family include the absence of an ITIM in Ly-49D, the lack of phosphorylation of Ly-49D in activated NK cells, and the association of a novel tyrosine-phosphorylated protein (pp16) with Ly-49D in activated NK cells
• Ly-49O and Ly-49V are closely related to Ly-49A[B6] and, like Ly-49A, have ITIM domains
• Ly-49O- and Ly-49V-transfected 293T (human kidney epithelial) cells bind tetramers of H-2D[b], D[b], D[k], and L[d]
• In addition, the Ly-49V-transfected cells also bind K[b], K[d], and K[k]
• Ly-49R is closely related to Ly-49D[B6] and is putative activating receptor due to its lack of an ITIM domain
• Ly-49R-transfected 293T cells bind soluble tetramers of H-2D[b], D[d], D[k], and L[d].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.