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      V450 Mouse Anti-Mouse Ly-49A
      V450 Mouse Anti-Mouse Ly-49A
      Flow cytometric analysis of Ly-49A expressed on C57BL/6 mouse splenocytes. C57BL/6 splenocytes were simultaneously stained with FITC Rat anti-Mouse CD49b (Cat. No. 553857) and BD Horizon™ V450 Mouse (BALB/c) anti-Mouse Ly-49A (Cat. No. 561204, Left Panel) or BD Horizon™ V450 Mouse IgG2a, κ Isotype Control (Cat. No. 560550, Right Panel). Two-color flow cytometric dot plots were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometry was performed on a BD LSR™ II Flow Cytometer System.
      V450 Mouse Anti-Mouse Ly-49A
      Flow cytometric analysis of Ly-49A expressed on C57BL/6 mouse splenocytes. C57BL/6 splenocytes were simultaneously stained with FITC Rat anti-Mouse CD49b (Cat. No. 553857) and BD Horizon™ V450 Mouse (BALB/c) anti-Mouse Ly-49A (Cat. No. 561204, Left Panel) or BD Horizon™ V450 Mouse IgG2a, κ Isotype Control (Cat. No. 560550, Right Panel). Two-color flow cytometric dot plots were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometry was performed on a BD LSR™ II Flow Cytometer System.
      Product Details
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      BD Horizon™
      Ly-49a; Klra1; Killer cell lectin-like receptor subfamily A member1; Klra22
      Mouse (QC Testing)
      Mouse BALB/c IgG2a, κ
      Mouse C57BL/6N T lymphoma EL-4
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      16627
      AB_10611583
      Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon™ V450 under optimum conditions, and unreacted BD Horizon™ V450 was removed.
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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. An isotype control should be used at the same concentration as the antibody of interest.
      3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
      4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
      6. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
      7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      561204 Rev. 2
      Antibody Details
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      A1

      The A1 monoclonal antibody specifically binds to the Ly-49A[B6] alloantigen, an inhibitory receptor that is expressed on subsets of natural killer (NK) cells and NK-1.1-positive T lymphocytes (NKT cells) in C57BL/6, C57BL/10, and B10 congenic mice, on a population of memory CD8+ T lymphocytes and NK1.1+ γδ T cells in C57BL/6 mice, and on a distinct subset of B-1 cells (CD5+B220[lo]) of C57BL/6 mice. The A1 antibody has also been reported to crossreact with Ly-49ANOD, Ly-49PNOD, Ly-49P129/J, and Ly-49V129/J alloantigens. The proportion of NKT cells expressing Ly-49A is higher (2-5 fold) in thymus than in liver (immature and mature NKT cells, respectively), and there is evidence that the down regulation of Ly-49 receptor expression is necessary for normal NKT cell development to occur. Most NK cells express a single allele of Ly-49A, although occasionally they may express more than one allele. 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 (CRD) that bind to MHC class I alloantigens. The A1 antibody is specific for the Ly-49A[B6] CRD. 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 Ly-49A[B6] allonantigen binds to H-2D[d], H-2D[k], and H-2D[p], and the A1 antibody blocks this binding. Binding of Ly-49A[B6] to lyphoblasts expressing MHC class I antigens of the f, q, r, s, and v haplotypes has also been demonstrated. The levels of the Ly-49 inhibitory receptors are down-regulated by their ligands in vivo, and various levels of expression of a Ly-49 inhibitory receptor may affect the specificity of NK cells. In vitro studies suggest that the Ly-49A receptor mediates negative regulation of NK-cell cytolytic activity via tyrosine phosphorylation of its ITIM (Immunoreceptor Tyrosine-based Inhibitory Motif).  

      The antibody is conjugated to BD Horizon™ V450, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD Horizon™ V450 can be used in place of Pacific Blue™ conjugates.

      561204 Rev. 2
      Format Details
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      V450
      BD Horizon™ V450 Dye is part of the BD Horizon™ violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 405-nm and an emission maximum (Em Max) at 450-nm. BD Horizon™ V450, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      V450
      Violet 405 nm
      405 nm
      450 nm
      561204 Rev.2
      Citations & References
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      Development References (32)

      1. Bendelac A. Mouse NK1+ T cells. Curr Opin Immunol. 1995; 7(3):367-374. (Clone-specific). View Reference
      2. Brennan J, Mahon G, Mager DL, Jefferies WA, Takei F. Recognition of class I major histocompatibility complex molecules by Ly-49: specificities and domain interactions. J Exp Med. 1996; 183(4):1553-1559. (Biology). View Reference
      3. Chang CS, Kane KP. Evidence for sulfate modification of H-2Dd on N-linked carbohydrate(s): possible involvement in Ly-49A interaction. J Immunol. 1998; 160(9):4367-4374. (Biology). View Reference
      4. Coles MC, McMahon CW, Takizawa H, Raulet DH. Memory CD8 T lymphocytes express inhibitory MHC-specific Ly49 receptors. Eur J Immunol. 2000; 30(1):236-244. (Biology). View Reference
      5. Daniels BF, Karlhofer FM, Seaman WE, Yokoyama WM. A natural killer cell receptor specific for a major histocompatibility complex class I molecule. J Exp Med. 1994; 180(2):687-692. (Biology). View Reference
      6. Guy-Grand D, Cuenod-Jabri B, Malassis-Seris M, Selz F, Vassalli P. Complexity of the mouse gut T cell immune system: identification of two distinct natural killer T cell intraepithelial lineages. Eur J Immunol. 1996; 26(9):2248-2256. (Clone-specific). View Reference
      7. Hanke T, Takizawa H, McMahon CW, et al. Direct assessment of MHC class I binding by seven Ly49 inhibitory NK cell receptors. Immunity. 1999; 11(1):67-77. (Biology). View Reference
      8. Hara T, Nishimura H, Hasegawa Y, Yoshikai Y. Thymus-dependent modulation of Ly49 inhibitory receptor expression on NK1.1+gamma/delta T cells. Immunology. 2001; 102(1):24-30. (Biology). View Reference
      9. Held W, Kunz B. An allele-specific, stochastic gene expression process controls the expression of multiple Ly49 family genes and generates a diverse, MHC-specific NK cell receptor repertoire. Eur J Immunol. 1998; 28(8):2407-2416. (Biology). View Reference
      10. Held W, Roland J, Raulet DH. Allelic exclusion of Ly49-family genes encoding class I MHC-specific receptors on NK cells. Nature. 1995 July; 376(6538):355-358. (Biology). View Reference
      11. Hoglund P, Sundback J, Olsson-Alheim MY, et al. Host MHC class I gene control of NK-cell specificity in the mouse. Immunol Rev. 1997; 155:11-28. (Biology). View Reference
      12. Kane KP. Ly-49 mediates EL4 lymphoma adhesion to isolated class I major histocompatibility complex molecules. J Exp Med. 1994; 179(3):1011-1015. (Biology). View Reference
      13. Karlhofer FM, Ribaudo RK, Yokoyama WM. MHC class I alloantigen specificity of Ly-49+ IL-2-activated natural killer cells. Nature. 1992; 358(6381):66-70. (Clone-specific). View Reference
      14. Kase A, Johansson MH, Olsson-Alheim MY, Karre K, Hoglund P. External and internal calibration of the MHC class I-specific receptor Ly49A on murine natural killer cells. J Immunol. 1998; 161(11):6133-6138. (Biology). View Reference
      15. Kim S, Yokoyama WM. NK cell granule exocytosis and cytokine production inhibited by Ly-49A engagement. Cell Immunol. 1998; 183(2):106-112. (Biology). View Reference
      16. Liu J, Yu YY, Lindahl KF, Kumar V, Bennett M. Allorecognition by murine natural killer cells: studies with bone marrow transplants and lysis of T lymphoblasts. Chem Immunol. 1996; 64:164-180. (Biology). View Reference
      17. Makrigiannis AP, Pau AT, Saleh A, Winkler-Pickett R, Ortaldo JR, Anderson SK. Class I MHC-binding characteristics of the 129/J Ly49 repertoire. J Immunol. 2001; 166(8):5034-5043. (Biology). View Reference
      18. 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). View Reference
      19. Mason LH, Willette-Brown J, Anderson SK, et al. Characterization of an associated 16-kDa tyrosine phosphoprotein required for Ly-49D signal transduction. J Immunol. 1998; 160(9):4148-4152. (Biology). View Reference
      20. Matsumoto N, Ribaudo RK, Abastado JP, Margulies DH, Yokoyama WM. The lectin-like NK cell receptor Ly-49A recognizes a carbohydrate-independent epitope on its MHC class I ligand. Immunity. 1998; 8(2):245-254. (Biology). View Reference
      21. 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. (Immunogen: Cytotoxicity, Flow cytometry, Immunoprecipitation). View Reference
      22. Ochi H, Watanabe T. Negative regulation of B cell receptor-mediated signaling in B-1 cells through CD5 and Ly49 co-receptors via Lyn kinase activity. Int Immunol. 2000; 12(10):1417-1423. (Biology). View Reference
      23. Olsson-Alheim MY, Salcedo M, Ljunggren HG, Karre K, Sentman CL. NK cell receptor calibration: effects of MHC class I induction on killing by Ly49Ahigh and Ly49Alow NK cells. J Immunol. 1997; 159(7):3189-3194. (Biology). View Reference
      24. Olsson-Alheim MY, Sundback J, Karre K, Sentman CL. The MHC class I molecule H-2Dp inhibits murine NK cells via the inhibitory receptor Ly49A. J Immunol. 1999; 162(12):7010-7014. (Biology). View Reference
      25. Raulet DH, Held W, Correa I, Dorfman JR, Wu MF, Corral L. Specificity, tolerance and developmental regulation of natural killer cells defined by expression of class I-specific Ly49 receptors. Immunol Rev. 1997; 155:41-52. (Biology). View Reference
      26. Robson MacDonald H, Lees RK, Held W. Developmentally regulated extinction of Ly-49 receptor expression permits maturation and selection of NK1.1+ T cells. J Exp Med. 1998; 187(12):2109-2114. (Biology). View Reference
      27. Silver ET, Gong DE, Chang CS, Amrani A, Santamaria P, Kane KP. Ly-49P activates NK-mediated lysis by recognizing H-2Dd. J Immunol. 2000; 165(4):1771-1781. (Biology). View Reference
      28. Skold M, Cardell S. Differential regulation of Ly49 expression on CD4+ and CD4-CD8- (double negative) NK1.1+ T cells. Eur J Immunol. 2000; 30(9):2488-2496. (Clone-specific). View Reference
      29. Sundback J, Nakamura MC, Waldenstrom M, et al. The alpha2 domain of H-2Dd restricts the allelic specificity of the murine NK cell inhibitory receptor Ly-49A. J Immunol. 1998; 160(12):5971-5978. (Biology). View Reference
      30. Yokoyama WM, Kehn PJ, Cohen DI, Shevach EM. Chromosomal location of the Ly-49 (A1, YE1/48) multigene family. Genetic association with the NK 1.1 antigen. J Immunol. 1990; 145(7):2353-2358. (Clone-specific: Flow cytometry). View Reference
      31. Yokoyama WM, Seaman WE. The Ly-49 and NKR-P1 gene families encoding lectin-like receptors on natural killer cells: the NK gene complex. Annu Rev Immunol. 1993; 11:613-635. (Clone-specific). View Reference
      32. Yu YY, George T, Dorfman JR, Roland J, Kumar V, Bennett M. The role of Ly49A and 5E6(Ly49C) molecules in hybrid resistance mediated by murine natural killer cells against normal T cell blasts. Immunity. 1996; 4(1):67-76. (Biology). View Reference
      View All (32) View Less
      561204 Rev. 2

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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.

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