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Put all BD® AbSeq Reagents to be pooled into a Latch Rack for 500 µL Tubes (Thermo Fisher Scientific Cat. No. 4900). Arrange the tubes so that they can be easily uncapped and re-capped with an 8-Channel Screw Cap Tube Capper (Thermo Fisher Scientific Cat. No. 4105MAT) and the reagents aliquoted with a multi-channel pipette.
BD® AbSeq tubes should be centrifuged for ≥ 30 seconds at 400 × g to ensure removal of any content in the cap/tube threads prior to the first opening.
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- This reagent has been pre-diluted for use at the recommended volume per test. Typical use is 2 µl for 1 × 10^6 cells in a 200-µl staining reaction.
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コンパニオン製品
The 5E6 (also known as clone SW5E6) antibody reacts with Ly-49C[BALB], Ly-49C[B6], Ly-49C[NZB], and Ly-49I[B6], inhibitory receptors which are expressed on subsets of natural killer (NK) cells and NK-1.1+ (or DX5+) T lymphocytes (NK-T cells) in all strains tested except C57BR and RIII, 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 of BALB/c and C57BL/6 mice. The proportion of NK T cells expressing Ly-49C/I is higher (2-5 fold) in thymus than in liver (immature and mature NK T cells, respectively), and there is evidence that the down-regulation of Ly-49 receptor expression is necessary for normal NK T-cell development. Most NK cells express a single allele of Ly-49C, although occasionally they may express more than one allele. The Ly-49 family of NK-cell receptors are disulfide-linked type-II transmembrane protein homodimers with extracellular carbohydrate-recognition domains (CRD) that 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. The 5E6 antibody is specific for the Ly-49C CRD. The Ly-49C[BALB] and Ly-49C[B6] alloantigens bind to MHC class I antigens of the b, d, k, and s haplotypes, and the 5E6 antibody blocks this binding. Binding of Ly-49C[BALB]- and Ly-49C[B6]- expressing transfectants to lymphoblasts of H-2[f], H-2[q], H-2[r], and H-2[v] strains has also been detected. Ly-49I[B6] transfectants bind H-2[r] lymphoblasts and bind much more weakly to the b, d, k, q, s, and v haplotypes. The levels of the Ly-49 inhibitory receptors are down-regulated by their ligands in vivo, and the various levels of expression of an Ly-49 inhibitory receptor may affect the specificity of NK cells. Ly-49C is specifically downregulated in the presence of H-2K[b] class I molecules (one of the Ly-49C ligands). Ly-49C[+] and/or Ly-49I[+] cells mediate allogeneic and hybrid resistance to H-2d bone marrow transplantation. In vitro and in vivo studies suggest that the Ly-49C and/or Ly-49I receptors mediate negative regulation of NK-cell cytolytic activity via tyrosine phosphorylation of their ITIMs (Immunoreceptor Tyrosine-based Inhibitory Motifs).
The epitope recognized by this antibody on Ly49C may be masked on freshly isolated primary NK cells due to cis interactions with MHC class I molecules. This observation has been reported for other Ly49C monoclonal antibodies that bind to the same structural region.
開発者向け参考資料 (24)
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Brennan J, Lemieux S, Freeman JD, Mager DL, Takei F. Heterogeneity among Ly-49C natural killer (NK) cells: characterization of highly related receptors with differing functions and expression patterns. J Exp Med. 1996; 184(6):2085-2090. (Biology). 参考文献を見る
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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). 参考文献を見る
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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). 参考文献を見る
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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). 参考文献を見る
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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). 参考文献を見る
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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). 参考文献を見る
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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). 参考文献を見る
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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). 参考文献を見る
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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). 参考文献を見る
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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). 参考文献を見る
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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). 参考文献を見る
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Ortaldo JR, Winkler-Pickett R, Mason AT, Mason LH. The Ly-49 family: regulation of cytotoxicity and cytokine production in murine CD3+ cells. J Immunol. 1998; 160(1):1158-1165. (Biology). 参考文献を見る
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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). 参考文献を見る
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Raziuddin A, Longo DL, Mason L, Ortaldo JR, Bennett M, Murphy WJ. Differential effects of the rejection of bone marrow allografts by the depletion of activating versus inhibiting Ly-49 natural killer cell subsets. J Immunol. 1998; 160(1):87-94. (Biology). 参考文献を見る
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Raziuddin A, Longo DL, Mason L, Ortaldo JR, Murphy WJ. Ly-49 G2+ NK cells are responsible for mediating the rejection of H-2b bone marrow allografts in mice. Int Immunol. 1996; 8(12):1833-1839. (Biology). 参考文献を見る
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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). 参考文献を見る
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Salcedo M, Andersson M, Lemieux S, Van Kaer L, Chambers BJ, Ljunggren HG. Fine tuning of natural killer cell specificity and maintenance of self tolerance in MHC class I-deficient mice. Eur J Immunol. 1998; 28(4):1315-1321. (Biology). 参考文献を見る
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Scarpellino L, Oeschger F, Guillaume P, et al. Interactions of Ly49 family receptors with MHC class I ligands in trans and cis.. J Immunol. 2007; 178(3):1277-1284. (Biology). 参考文献を見る
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Sentman CL, Hackett J, Kumar V, Bennett M. Identification of a subset of murine natural killer cells that mediates rejection of Hh-1d but not Hh-1b bone marrow grafts. J Exp Med. 1989; 170(1):191-202. (Immunogen). 参考文献を見る
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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. (Biology). 参考文献を見る
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Stoneman ER, Bennett M, An J, et al. Cloning and characterization of 5E6(Ly-49C), a receptor molecule expressed on a subset of murine natural killer cells. J Exp Med. 1995; 182(2):305-313. (Biology). 参考文献を見る
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Takei F, Brennan J, Mager DL. The Ly-49 family: genes, proteins and recognition of class I MHC. Immunol Rev. 1997; 155:67-77. (Biology). 参考文献を見る
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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). 参考文献を見る
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Yu YY, Kumar V, Bennett M. Murine natural killer cells and marrow graft rejection. Annu Rev Immunol. 1992; 10:189-213. (Biology). 参考文献を見る
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