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Purified Rat Anti-Mouse Ly-49G2
Purified Rat Anti-Mouse Ly-49G2

Two-color analysis of Ly-49G2 expression on splenic NK cells. C57BL/6 splenocytes were simultaneously stained with PE-conjugated anti-mouse NK-1.1 mAb PK136 (Cat. No. 557391/553165, both panels) and purified 4D11 monoclonal antibody (right panel), followed by FITC conjugated goat anti-rat Ig (Cat. No. 554001, both panels) and purified 4D11 monoclonal antibody (right panel), followed by FITC-conjugated goat anti-rat Ig (Cat. No. 554016, both panels). Flow cytometry was performed on a BD FACScan™ Flow Cytometry System.

Two-color analysis of Ly-49G2 expression on splenic NK cells. C57BL/6 splenocytes were simultaneously stained with PE-conjugated anti-mouse NK-1.1 mAb PK136 (Cat. No. 557391/553165, both panels) and purified 4D11 monoclonal antibody (right panel), followed by FITC conjugated goat anti-rat Ig (Cat. No. 554001, both panels) and purified 4D11 monoclonal antibody (right panel), followed by FITC-conjugated goat anti-rat Ig (Cat. No. 554016, both panels). Flow cytometry was performed on a BD FACScan™ Flow Cytometry System.

Product Details
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BD Pharmingen™
LGL-1; Klra7
Mouse (QC Testing)
Rat F344, also known as Fischer, CDF IgG2a, κ
Large granular lymphocytes (LGL) enriched from C57BL/6N mouse liver
Flow cytometry (Routinely Tested), Activation, Blocking, Cytotoxicity, Depletion, Immunoprecipitation (Reported)
0.5 mg/ml
16638
AB_395725
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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.
555314 Rev. 8
Antibody Details
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4D11

The 4D11 antibody specifically recognizes Ly-49G2 (also known as LGL-1), an inhibitory receptor which is expressed on subsets of natural killer (NK) cells and DX5-positive T lymphocytes (NK-T cells) in all strains tested (e.g., AKR/N, BALB/c, C3H/HeJ, C57BL/6, CBA/J, DBA/2, SJL, 129) and on a population of memory CD8+ T lymphocytes in C57BL/6 mice. Cross-reaction of 4D11 antibody to Ly-49A[B6], Ly-49A[BALB], and Ly-49T[129/J] inhibitory receptors and Ly-49L[CBA/J] activating receptor has been reported. The proportion of NK-T cells expressing Ly-49A and Ly-49G2 is higher (2-5 fold) in thymus than in liver (immature and mature NK-T cells, respectively), and there is evidence that down-regulation of Ly-49 receptor expression is necessary for normal NK-T-cell development to occur. Most NK cells express a single allele of Ly-49A and/or Ly-49G2, 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, which 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. Binding of Ly-49G[B6]-expressing transfectants to H-2Dd+/H-2Ld+ ConA blasts has been demonstrated, and H-2D[d]-expressing target cells inhibit the lytic activity of Ly-49G2-expressing NK cells. The levels of the Ly-49 inhibitory receptors are down-regulated by their ligands in vivo, and the various levels of expression of a Ly-49 inhibitory receptor may affect the specificity of NK cells. Ly-49G2[+] NK cells are able to lyse target tumor cells expressing H-2[a] and H-2[b] MHC class I antigens in vitro, and they mediate allogeneic and hybrid resistance to H-2[b] bone marrow transplantation. The Ly-49A[BALB] and Ly-49A[B6] alloantigens bind to MHC class I antigens of the d and k haplotypes, and Ly-49A[+] IL-2-activated NK cells are unable to lyse target cells expressing H-2[d] and H-2[k]. In vitro studies suggest that the Ly-49G2 and Ly-49A receptors mediate negative regulation of NK-cell cytolytic activity via tyrosine phosphorylation of their ITIMs (Immunoreceptor Tyrosine-based Inhibitory Motifs). Ly-49T[129/J] has a unique ITIM sequence, and Ly-49T-transfected 293T (human kidney epithelial) cells do not bind soluble tetramers of any tested H-2 alloantigen (D[b], D[d], D[k], K[b], K[d], K[k], L[d]).

555314 Rev. 8
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
555314 Rev.8
Citations & References
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Development References (20)

  1. 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
  2. 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
  3. 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
  4. 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
  5. Johansson MH, Höglund E, Nakamura MC, Ryan JC, Höglund P. Alpha1/alpha2 domains of H-2D(d), but not H-2L(d), induce "missing self" reactivity in vivo--no effect of H-2L(d) on protection against NK cells expressing the inhibitory receptor Ly49G2. Eur J Immunol. 1998; 28(12):4198-4206. (Clone-specific). View Reference
  6. Makrigiannis AP, Etzler J, Winkler-Pickett R, Mason A, Ortaldo JR, Anderson SK. Identification of the Ly49L protein: evidence for activating counterparts to inhibitory Ly49 proteins. J Leukoc Biol. 2000; 68(5):765-771. (Biology). View Reference
  7. 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
  8. Mason L, Giardina SL, Hecht T, Ortaldo J, Mathieson BJ. LGL-1: a non-polymorphic antigen expressed on a major population of mouse natural killer cells. J Immunol. 1988; 140(12):4403-4412. (Immunogen: Immunoprecipitation). View Reference
  9. 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. (Clone-specific). View Reference
  10. Mason LH, Ortaldo JR, Young HA, Kumar V, Bennett M, Anderson SK. Cloning and functional characteristics of murine large granular lymphocyte-1: a member of the Ly-49 gene family (Ly-49G2). J Exp Med. 1995; 182(2):293-303. (Clone-specific: Blocking, Immunoprecipitation). View Reference
  11. Mason LH, Yagita H, Ortaldo JR. LGL-1: a potential triggering molecule on murine NK cells. J Leukoc Biol. 1994; 55(3):362-370. (Clone-specific: Mediation). View Reference
  12. 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
  13. Ortaldo JR, Mason AT, Winkler-Pickett R, Raziuddin A, Murphy WJ, Mason LH. Ly-49 receptor expression and functional analysis in multiple mouse strains. J Leukoc Biol. 1999; 66(3):512-520. (Clone-specific: Blocking). View Reference
  14. 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). View Reference
  15. 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
  16. 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. (Clone-specific). View Reference
  17. 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. (Clone-specific). View Reference
  18. 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
  19. 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). View Reference
  20. 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). View Reference
View All (20) View Less
555314 Rev. 8

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