Phosphate-buffered solution, pH 7.2, containing no preservative. 0.2 μm filter sterilized. Endotoxin level is < 0.1 EU/μg of the protein (< 0.01 ng/μg of the protein) as determined by the LAL test.
Preparation:
The antibody was purified by affinity chromatography.
Concentration:
1 mg/ml
Storage & Handling:
The antibody solution should be stored undiluted at 4 °C. This LEAF™ solution contains no preservative; handle under aseptic conditions.
Application:
FC - Quality tested Blocking1-5*, IHC(Paraffin)6* - *This application has been reported in the literature.
Recommended Usage:
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is ≤ 0.25 µg per 106 cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes:
Additional reported applications (for the relevant formats) include: in vitro blocking1,2 and in vivo blocking3-5, and immunohistochemical staining6 of formalin-fixed paraffin-embedded tissue sections. Fas Ligand is expressed at low density on activated cells. For most successful immunofluorescent staining results, it may be important to maximize signal over background by using a relatively bright fluorochrome-antibody conjugate (Cat. No. 106806) or by using a high sensitivity, three-layer staining technique (e.g., including a biotinylated antibody (Cat. No. 106804) or biotinylated anti-mouse IgG (Cat. No. 405303) second step , followed by SAv-PE (Cat. No. 405204)). The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for functional assays (Cat. No. 106808).
Application References:
1. Kayagaki, N., et al., 1997. PNAS 94:3914. 2. Kobata, T., et al., 1997. Immunology 92:206. 3. Hattori, K., et al., 1998. Blood 91:4051. 4. Takeda, Y., et al., 1998. Diabetologia 41:315. 5. Kim, Y.-H., et al., 1999. Eur. J. Immunol. 29:455. 6. Takeda, Y., et al., 1998. Diabetologia 41:315.
Mouse Fas Ligand transfected cells stained with LEAF™ purified KAY-10, followed by biotinylated anti-mouse IgG and Sav-PE
Description:
CD178.1 is a 40 kD Fas ligand alloantigen (designated as mFasL.1) expressed on the activated T lymphocytes, testis, and eye of selected mouse strains including C57BL/6, C3H, MRL, NOD, NZB, NZW, and SJL. It does not react with lymphoblasts from BALB/c, DBA/1, or DBA/2 mice. CD178.1 is upregulated in activated T cells upon TCR re-engagement and has been shown to induce autocrine and paracrine T cell death. CD178 expression in the eye and testis has been shown to participate in immune privilege at these sites. Functional studies suggest that mFasL.2 has higher specific activity than mFasL.1. CD178 can be cleaved from the surface by metalloproteases, and "soluble" CD178 may block the activities of membrane-bound CD178. CD178 binds to CD95 (Fas) to induce apoptotic cell death implicated in the maintenance of peripheral tolerance. CD178/CD95 interactions have been implicated in the proliferation of CD8+ cells and neutrophil extravasation, chemotaxis, and survival. The Kay-10 antibody has been reported to block CD178.1/CD95 induced apoptosis in cells derived from mice expressing this alloantigen.
Other Names:
Fas Ligand, FasL, Apo-1 Ligand, CD95 Ligand
Structure:
TNF superfamily Fas ligand alloantigen, 40 kD
Distribution:
Activated T cells, spleen, testis, eye of some selected mouse strains (C57BL/6, C3H, NZB, NZw, MRL)
Function:
Induces apoptosis
Ligand Receptor:
Fas (CD95)
Antigen References:
1. Barclay, A., et al., 1997. The Leukocyte Antigen FactsBook, Academic Press. 2. Nagata, S. 1999. Annu. Rev. Genet. 33:209. 3. Takahashi, T., et al., 1994. Cell 76:969. 4. Hill, L.L., et al., 1999. Science 285:898. 5. Kayagaki, N., et al., 1997. PNAS 94:3914.
