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PE Mouse Anti-Mouse Ly-49A
PE Mouse Anti-Mouse Ly-49A

Two-color analysis of Ly-49A expression on splenic NK cells. C57BL/6 splenocytes were simultaneously stained with FITC Rat Anti-Mouse CD49b (Cat. No. 553857) and PE Mouse Anti-Mouse Ly-49A (Cat. No. 557424; right panel). Two-color contour plots were derived from gated events with the side and forward side-scattering characteristics of viable splenocytes. Flow cytometry was performed on a BD FACScan™ flow cytometry system.

Two-color analysis of Ly-49A expression on splenic NK cells. C57BL/6 splenocytes were simultaneously stained with FITC Rat Anti-Mouse CD49b (Cat. No. 553857) and PE Mouse Anti-Mouse Ly-49A (Cat. No. 557424; right panel). Two-color contour plots were derived from gated events with the side and forward side-scattering characteristics of viable splenocytes. Flow cytometry was performed on a BD FACScan™ flow cytometry system.

Product Details
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BD Pharmingen™
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_396696
Aqueous buffered solution containing ≤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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
557424 Rev. 15
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).  

557424 Rev. 15
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
557424 Rev.15
Citations & References
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Development References (31)

  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. 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
  11. 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
  12. 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
  13. 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
  14. 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
  15. 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
  16. 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
  17. 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
  18. 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
  19. 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
  20. 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). View Reference
  21. 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
  22. 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
  23. 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
  24. 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
  25. 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
  26. 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
  27. 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
  28. 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
  29. 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). View Reference
  30. 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
  31. 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 (31) View Less
557424 Rev. 15

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.