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Multiparameter flow cytometric analysis using BD OptiBuild™ BV421 Mouse Anti-Mouse Ly-49H antibody (Cat. No. 744260) on live BALB/c mouse splenocytes (left panel) and live C57BL/6 mouse splenocytes (right panel). Flow cytometry was performed using a BD LSRFortessa™ Flow Cytometer System.
BD OptiBuild™ BV421 Mouse Anti-Mouse Ly-49H
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Preparation And Storage
Recommended Assay Procedures
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For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
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The 3D10 monoclonal antibody specifically binds to mouse Lymphocyte antigen 49H (Ly-49H; also known as Klra8 or Killer cell lectin-like receptor 8). The 3D10 antibody does not crossreact with related molecules such as Ly-49A, C, D or G2. Ly-49H is a type II transmembrane protein and a member of the Ly-49 C-type lectin multigene family of receptors expressed by NK cells. Cell surface Ly-49H is expressed by a subset of NK cells but not by NKT cells. Ly-49H is expressed by C57BL/6 and NWNA but not by BALB/c or DBA/2 mouse NK cells. Cell surface Ly-49H presents as a ~110 kDa disfulfide-linked homodimer and associates with signaling subunits such as DAP10 and DAP12 for optimal transduction of intracellular activation signals. Crosslinking of Ly-49H with the 3D10 antibody reportedly induces NK cell cytotoxicity and cytokine production. Ly-49H recognizes the mouse cytomegalovirus m157 glycoprotein that is expressed by infected cells and is required for protection against cytomegalovirus infection.
Development References (5)
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Brennan J, Mager D, Jefferies W, Takei F. Expression of different members of the Ly-49 gene family defines distinct natural killer cell subsets and cell adhesion properties. J Exp Med. 1994; 180(6):2287-2295. (Biology). View Reference
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Brown MG, Dokun AO, Heusel JW, et al. Vital involvement of a natural cell activation receptor in resistance to viral infection. Science. 2001; 292(5518):934-937. (Clone-specific: Activation, Bioassay, Blocking, Cytotoxicity, Flow cytometry). View Reference
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Orr MT, Sun JC, Hesslein DG, et al. Ly49H signaling through DAP10 is essential for optimal natural killer cell responses to mouse cytomegalovirus infection. J Exp Med. 2009; 206(4):807-817. (Clone-specific: Blocking, Flow cytometry). View Reference
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Silver ET, Elliott JF, Kane KP. Alternatively spliced Ly-49D and H transcripts are found in IL-2-activated NK cells. Immunogenetics. 1996; 44(6):478-482. (Biology). View Reference
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Smith HR, Chuang HH, Wang LL, Salcedo M, Heusel JW, Yokoyama WM. Nonstochastic Coexpression of activation receptors on murine Natural Killer cells. J Exp Med. 2000; 191(8):1341-1354. (Immunogen: Activation, Cytotoxicity, Flow cytometry, Immunoprecipitation). View Reference
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.