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    PE Hamster Anti-Mouse CD178
    PE Hamster Anti-Mouse CD178
    Expression of Fas Ligand on T lymphocytes. T lymphocytes from C57BL/6 spleen (mouse T Cell Enrichment Column, R&D Systems, Minneapolis, MN), cultured for 7 hours on plate-bound anti-mouse CD3e mAb 500A2 (Cat. No. 553238, top panels) or on uncoated plates (bottom panels), were simultaneously stained with FITC-conjugated anti-mouse CD4 mAb RM4-5 (Cat. No. 553046/553047, top and bottom, left and center panels) and PE-conjugated mAb MFL3 (middle panels). Staining with PE-conjugated anti-mouse CD69 mAb H1.2F3 demonstrates the activation state of the T cells (Cat. No. 553237, right panels). Flow cytometry was performed on a BD FACScan™ flow cytometry system.
    PE Hamster Anti-Mouse CD178
    Expression of Fas Ligand on T lymphocytes. T lymphocytes from C57BL/6 spleen (mouse T Cell Enrichment Column, R&D Systems, Minneapolis, MN), cultured for 7 hours on plate-bound anti-mouse CD3e mAb 500A2 (Cat. No. 553238, top panels) or on uncoated plates (bottom panels), were simultaneously stained with FITC-conjugated anti-mouse CD4 mAb RM4-5 (Cat. No. 553046/553047, top and bottom, left and center panels) and PE-conjugated mAb MFL3 (middle panels). Staining with PE-conjugated anti-mouse CD69 mAb H1.2F3 demonstrates the activation state of the T cells (Cat. No. 553237, right panels). Flow cytometry was performed on a BD FACScan™ flow cytometry system.
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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)
    0.2 mg/ml
    AB_395711
    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. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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    Recommended Assay Procedures

    We have found that enriched splenic T cells are 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. 557306/553058), or mAb 500A2 (Cat. No. 553238).

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    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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
    4. 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.
    5. 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.
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    555293 Rev. 9
    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.

    555293 Rev. 9
    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
    555293 Rev.9
    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: Inhibition). 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: Inhibition). 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
    View All (17) View Less
    555293 Rev. 9

    Please refer to Support Documents for Quality Certificates


    Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


    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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