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Purified NA/LE Hamster Anti-Mouse CD178
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BD Pharmingen™
Fas Ligand, FasL; Fas-Lg; CD95 Ligand; CD95L; Tnfsf6; APT1LG1; gld
Mouse (QC Testing)
Armenian Hamster IgG1, κ
Mouse FasL-transfected cells
Flow cytometry (Routinely Tested), Blocking (Reported)
1.0 mg/ml
AB_395708
No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2µm sterile filtered. Endotoxin level is ≤0.01 EU/µg (≤0.001 ng/µg) of protein as determined by the LAL assay.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. This preparation contains no preservatives, thus it should be handled under aseptic conditions.

Recommended Assay Procedures

Enriched splenic T cells can be induced to express Fas Ligand by 6-8-hour culture with plate-bound anti-mouse CD3 mAb 17A2 (Cat. No. 555273), mAb 145-2C11 (Cat. No. 557306), or mAb 500A2 (Cat. No. 553238). Because Fas Ligand is expressed at low density on activated cells, it may be desirable to amplify staining by using a biotinylated second-step antibody with a "bright" third-step reagent, such as PE Streptavidin (Cat. No. 554061). The BD Biosciences anti-hamster IgG mAb cocktails, either biotin- or PE-conjugated (Cat. No. 554010 or 554056, respectively) are not effective for staining with mAb MFL3. Other reported applications include blocking of the cytotoxic activities of the mouse FasLigand-transfected L5178Y T lymphoma and influenza-specific CD8+ BALB/c CTL clones.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
555290 Rev. 10
Antibody Details
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MFL3

The MFL3 monoclonal antibody specifically binds to CD178 (Fas Ligand, CD95 Ligand) on all strains tested. In the mouse, Fas Ligand is expressed on activated T cell lines and in spleen, testis, and eye. FasL mRNA has been demonstrated at various levels in bone marrow, thymus, spleen, lymph node, lung, small intestine, testis, and uterus. Moreover, T-cell activators, but not B-cell activators, enhanced the expression of FasL mRNA in splenocytes; and FasL mRNA was restricted to the T-cell lineage among a panel of cell lines from lymphoid tissues. Fas Ligand is not functional in mice homozygous for the gld (generalized lympho-proliferative disease) mutation; these mice cannot limit the expansion of activated lymphocytes and develop autoimmune disease. Fas Ligand is a member of the TNF/NGF family, which binds to CD95 (Fas), inducing apoptotic cell death. This Fas/Fas Ligand interaction is believed to participate in T-cell development, the regulation of immune responses, and cell-mediated cytotoxic mechanisms. There is mounting evidence that Fas Ligand is also proinflammatory, mediating neutrophil extravasation and chemotaxis. Fas Ligand is released from the surface of transfectant cells by metalloproteinases, and the soluble Fas Ligand may block the activities of the membrane-bound molecule. The MFL3 mAb has been  reported to efficiently inhibit the cytotoxicity of mouse Fas Ligand-transfected cells against human Fas-transfected cells. This hamster mAb to a mouse leukocyte antigen does not cross-react with rat leukocytes.

555290 Rev. 10
Format Details
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NA/LE
NA/LE refers to the culture and purification methods and buffer used to produce purified antibodies with no azide and low endotoxin: Aqueous buffered solution containing no preservative, 0.2µm sterile filtered. Endotoxin level is ≤0.01 EU/µg (≤0.001 ng/µg) of protein as determined by the LAL assay.NA/LE are perfectly suited to be used in culture or in vivo (for nonhuman studies) for functional assays — blocking, neutralizing, activation or depletion — where the presence of azide may damage cells or exogenous endotoxin may signal or activate cells.
NA/LE
555290 Rev.10
Citations & References
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Development References (17)

  1. Bellgrau D, Gold D, Selawry H, Moore J, Franzusoff A, Duke RC. A role for CD95 ligand in preventing graft rejection. Nature. 1995; 377(6550):630-632. (Biology). View Reference
  2. Brunner T, Mogil RJ, LaFace D, et al. Cell-autonomous Fas (CD95)/Fas-ligand interaction mediates activation-induced apoptosis in T-cell hybridomas. Nature. 1995; 373:441-444. (Biology). View Reference
  3. Fuller CL, Ravichandran KS, Braciale VL. Phosphatidylinositol 3-kinase-dependent and -independent cytolytic effector functions. J Immunol. 1999; 162(11):6337-6340. (Clone-specific: Blocking). View Reference
  4. Griffith TS, Brunner T, Fletcher SM, Green DR, Ferguson TA. Fas ligand-induced apoptosis as a mechanism of immune privilege. Science. 1995; 270(5239):1189-1192. (Biology). View Reference
  5. Griffith TS, Ferguson TA. The role of FasL-induced apoptosis in immune privilege. Immunol Today. 1997; 18(5):240-244. (Biology). View Reference
  6. Hohlbaum AM, Moe S, Marshak-Rothstein A. Opposing effects of transmembrane and soluble Fas ligand expression on inflammation and tumor cell survival. J Exp Med. 2000; 191(7):1209-1220. (Biology). View Reference
  7. Ju ST, Panka DJ, Cui H, et al. Fas(CD95)/FasL interactions required for programmed cell death after T-cell activation. Nature. 1995; 373(6513):444-448. (Biology). View Reference
  8. Kayagaki N, Yamaguchi N, Nagao F, et al. Polymorphism of murine Fas ligand that affects the biological activity.. Proc Natl Acad Sci USA. 1997; 94(8):3914-9. (Immunogen: Blocking). View Reference
  9. Kojima H, Shinohara N, Hanaoka S, et al. Two distinct pathways of specific killing revealed by perforin mutant cytotoxic T lymphocytes. Immunity. 1994; 1(5):357-364. (Biology). View Reference
  10. Lau HT, Yu M, Fontana A, Stoeckert CJ Jr. Prevention of islet allograft rejection with engineered myoblasts expressing FasL in mice. Science. 1996; 273(5271):109-112. (Biology). View Reference
  11. Lynch DH, Ramsdell F, Alderson MR. Fas and FasL in the homeostatic regulation of immune responses. Immunol Today. 1995; 16(12):569-574. (Biology). View Reference
  12. Ramsdell F, Seaman MS, Miller RE, Picha KS, Kennedy MK, Lynch DH. Differential ability of Th1 and Th2 T cells to express Fas ligand and to undergo activation-induced cell death. Int Immunol. 1994; 6(10):1545-1553. (Biology). View Reference
  13. Schneider P, Holler N, Bodmer JL, et al. Conversion of membrane-bound Fas(CD95) ligand to its soluble form is associated with downregulation of its proapoptotic activity and loss of liver toxicityq. J Exp Med. 1998; 187(8):1205-1213. (Biology). View Reference
  14. Smith CA, Farrah T, Goodwin RG. The TNF receptor superfamily of cellular and viral proteins: activation, costimulation, and death. Cell. 1994; 76(6):959-962. (Biology). View Reference
  15. Suda T, Okazaki T, Naito Y, et al. Expression of the Fas ligand in cells of T cell lineage. J Immunol. 1995; 154(8):3806-3813. (Biology). View Reference
  16. Takahashi T, Tanaka M, Brannan CI, et al. Generalized lymphoproliferative disease in mice, caused by a point mutation in the Fas ligand. Cell. 1994; 76(6):969-976. (Biology). View Reference
  17. Vignaux F, Vivier E, Malissen B, Depraetere V, Nagata S, Golstein P. TCR/CD3 coupling to Fas-based cytotoxicity. J Exp Med. 1995; 181(2):781-786. (Biology). View Reference
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555290 Rev. 10

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