Ly-49D Rat anti-Mouse, Brilliant Violet 605, 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].This antibody is conjugated to BD Horizon™ BV605 which is part of the BD Horizon Brilliant™ Violet family of dyes
• With an Ex Max of 407nm and Em Max of 602nm, BD Horizon BV605 can be excited by a violet laser and detected with a standard 610/20nm filter set
• BD Horizon BV605 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an Em max at 605nm
• Due to the excitation of the acceptor dye by the green (532 nm) and yellow-green (561 nm) lasers, there will be significant spillover into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers
• BD Horizon BV605 conjugates are very bright, often exhibiting brightness equivalent to PE conjugates and can be used as a third color off of the violet laser
• BV605 is a tandem fluorochrome that combines BD Horizon BV421 and Cy™3.5
• These conjugates are very bright, exhibiting similar brightness to equivalent PE conjugates
• Due to the excitation of Cy3.5 by the green (532nm) and yellow-green (561nm) lasers, there will be significant spillover of the BD Horizon BV605 signal into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers.