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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.
BD Pharmingen™ Biotin Mouse Anti-Mouse CD178.1
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준비 및 보관
권장 분석 절차
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.
제품 고시
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
관련 제품
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.
개발 참고 자료 (14)
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