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Flow cytometric analysis of Ly-49A expression on Mouse splenocytes. BALB/c Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with APC Rat Anti-Mouse CD49b antibody (Cat No. 560628) and with either PE Rat IgG2b, κ Isotype Control (Cat. No. 555848; Left Plot) or PE Mouse Anti-Rat Ly-49A antibody (Cat. No. 569156/569157; Right Plot) at 0.06 μg/test. 7-AAD (7-Amino-Actinomycin D) Solution (Cat. No. 559925) was added to cells right before analysis. A bivariate pseudocolor density plot showing the correlated expression of Ly-49A (or Ig Isotype control staining) versus CD49b was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ PE Rat Anti-Mouse Ly-49A
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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
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Companion Products
The YE1/48.10.6 monoclonal antibody specifically recognizes Lymphocyte antigen 49a (Ly-49A). Ly-49A is a single-pass type-II transmembrane that is encoded by Klra1 (Killer cell lectin-like receptor, subfamily A member 1) which belongs to the C-type lectin superfamily. Ly-49A is comprised of an extracellular carbohydrate-recognition domain (CRD) that can bind strongly to H-2D MHC class I alloantigens expressed on target cells including H-2D[d], H-2D[k], and H-2D[p]. The extracellular domain is followed by a transmembrane region and a cytoplasmic domain with an immunoreceptor tyrosine-based inhibitory motif (ITIM). Ly-49A is expressed as a disulfide-linked homodimer on a subset of NK and NK-T cells. MHC Class I ligand-bound Ly-49A can negatively regulate NK-cell cytolytic activity via tyrosine phosphorylation of its ITIM.
Development References (6)
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Chan PY, Takei F. Expression of a T cell receptor-like molecule on normal and malignant murine T cells detected by rat monoclonal antibodies to nonclonotypic determinants.. J Immunol. 1986; 136(4):1346-53. (Immunogen: Blocking, Flow cytometry, Immunoprecipitation, Western blot). View Reference
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Mason LH, Gosselin P, Anderson SK, Fogler WE, Ortaldo JR, McVicar DW. Differential tyrosine phosphorylation of inhibitory versus activating Ly-49 receptor proteins and their recruitment of SHP-1 phosphatase. J Immunol. 1997; 159(9):4187-4196. (Biology: Blocking). View Reference
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Nagasawa R, Gross J, Kanagawa O, et al. Identification of a novel T cell surface disulfide-bonded dimer distinct from the alpha/beta antigen receptor. J Immunol. 1987; 138(3):815-824. (Biology). View Reference
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Nakamura MC, Seaman WE. Ligand interactions by activating and inhibitory Ly-49 receptors.. Immunol Rev. 2001; 181:138-48. (Biology). View Reference
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Takei F. Two surface antigens expressed on proliferating mouse T lymphocytes defined by rat monoclonal antibodies.. J Immunol. 1983; 130(6):2794-7. (Immunogen: Flow cytometry, Radioimmunoassay, Western blot). View Reference
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Zimmer J, Ioannidis V, Held W. H-2D ligand expression by Ly49A+ natural killer (NK) cells precludes ligand uptake from environmental cells: implications for NK cell function.. J Exp Med. 2001; 194(10):1531-9. (Clone-specific: Flow cytometry). View Reference
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