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Biotin Mouse Anti-Mouse CD178.1
Biotin Mouse Anti-Mouse CD178.1
Expression of Fas Ligand on activated T lymphocytes. T lymphocytes from C57BL/6 spleen (mouse T Cell Enrichment Column, R&D Systems, Minneapolis, MN), cultured for 7 hours in the presence of plate-bound anti-mouse CD3e mAb 500A2 (Cat. no. 553238, upper panels) or on uncoated plates (lower panels were simultaneously stained with FITC-conjugated anti-mouse CD4 mAb RM4-5 (Cat. no. 553046/553047) and biotin-conjugated mAb Kay-10 (center panels) or PE-conjugated anti-mouse CD69 mAb H1.2F3 (to demonstrate activation, Cat. no. 553237, right panels), followed by Streptavidin-PE (Cat. no. 554061, left and center panels). Flow cytometry was performed on a BD FACScan™ flow cytometry system.
Expression of Fas Ligand on activated T lymphocytes. T lymphocytes from C57BL/6 spleen (mouse T Cell Enrichment Column, R&D Systems, Minneapolis, MN), cultured for 7 hours in the presence of plate-bound anti-mouse CD3e mAb 500A2 (Cat. no. 553238, upper panels) or on uncoated plates (lower panels were simultaneously stained with FITC-conjugated anti-mouse CD4 mAb RM4-5 (Cat. no. 553046/553047) and biotin-conjugated mAb Kay-10 (center panels) or PE-conjugated anti-mouse CD69 mAb H1.2F3 (to demonstrate activation, Cat. no. 553237, right panels), followed by Streptavidin-PE (Cat. no. 554061, left and center panels). Flow cytometry was performed on a BD FACScan™ flow cytometry system.
Product Details
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BD Pharmingen™
mFasL.1, Tnfsf6; CD95 Ligand; CD95L; Fasl; Faslg; gld;
Mouse (QC Testing)
Mouse C57BL/6 IgG2b, κ
Transfected Cell Line
Flow cytometry (Routinely Tested), Immunohistochemistry-formalin (antigen retrieval required) (Reported)
0.5 mg/ml
AB_395089
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.

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 antibody mAb 17A2 (Cat. no. 555273), 145-2C11 (Cat. no. 557306/553058), or 500A2 (Cat. no. 553238). Because Fas Ligand is expressed at low density on activated cells, we recommend the use of a "bright" second-step reagent, such as Streptavidin-PE (Cat. no. 554061). Other reported applications include immunohistochemical staining of formalin-fixed paraffin-embedded sections.

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.
553853 Rev. 11
Antibody Details
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KAY-10

The Kay-10 monoclonal antibody specifically recognizes CD178.1, the Fas ligand alloantigen (mFasL.1, CD95 ligand), which is expressed on activated T lymphocytes of selected strains of mice (eg, C57BL/6, C3H, MRL, NOD, NZB, NZW, and SJL). It does not react with similarly activated T blasts from BALB/c, DBA/1, or DBA/2 mice. It reacts with Cos cells transfected with mFasL cDNA derived from C57BL/6 and C3H mice, but not BALB/c or DBA/2 mice. The Kay-10 mAb efficiently blocks cytotoxic activity of a C3H T-cell line, but not of BALB/c. Phenotypic and genotypic characterizations of mouse Fas ligand reveal the existence of two alloantigens: mFasL.1, recognized by mAbs Kay-10, MFL3(Cat. no. 555291), and MFL4 (Cat. no. 555022); and mFasL.2, recognized by mAbs MFL3 and MFL4. Functional studies suggest that mFasL.2 has higher specific activity than mFasL.1. In the mouse, FasL is expressed on activated T cell lines and in the spleen, testis, and eye. FasL mRNA has been demonstrated at various levels in the 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; 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 lymphoproliferative 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 pro-inflammatory, mediating neutrophil extravasation and chemotaxis. Fas ligand is released from the surface of transfectant cells by metalloproteinases, and the soluble molecule may block the activities of the membrane-bound molecule.

553853 Rev. 11
Format Details
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Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
553853 Rev.11
Citations & References
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Development References (14)

  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. 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
  4. 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
  5. 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
  6. 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). View Reference
  7. 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
  8. 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
  9. Pestano GA, Zhou Y, Trimble LA, Daley J, Weber GF, Cantor H. Inactivation of misselected CD8 T cells by CD8 gene methylation and cell death. Science. 1999 May; 284(5417):1187-1191. (Biology). View Reference
  10. 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
  11. 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
  12. 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
  13. Takeda Y, Gotoh M, Dono K, et al. Protection of islet allografts transplanted together with Fas ligand expressing testicular allografts. Diabetologia. 1998 March; 41(3):315-321. (Clone-specific: Immunohistochemistry). View Reference
  14. 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 (14) View Less
553853 Rev. 11

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