L5178Y mouse T lymphoma cells expressing recombinant human FasL
Formulation:
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 IP1,2, Blocks Fas induced apoptosis1, IF3, ICC2* - *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 ≤ 1.0 µg per 106 cells in 100 µl volume or 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes:
Additional reported applications (for the relevant formats) include: immunoprecipitation1,2, immunofluorescence microscopy3, immunocytochemistry2, and blocking of Fas induced apoptosis1. 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. 306407) or by using a high sensitivity, three-layer staining technique (e.g., including a biotinylated antibody (Cat. No. 306404) or biotinylated anti-mouse IgG second step (Cat. No. 405303), followed by SAv-PE (Cat. No. 405204)). In addition, applying matrix metalloproteinases (MMPS) inhibitor in the cell culture system will increase the FasL staining intensity. The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for functional assays (Cat. No. 306409).
Application References:
1. Kayagaki, N., et al., 1995. J. Exp. Med. 182:1777. 2. Andreola, G., et al., 2002. J. Exp. Med. 195:1303. 3. Bossi, G., et al., 1999. Nature Medicine 5:90.
Human Fas Ligand transfected cells stained with LEAF™ purified NOK-1, followed by biotinylated anti-mouse IgG and Sav-PE
Description:
CD178 is a 38-42 kD type II glycoprotein also known as Fas ligand and CD95L. CD178 belongs to the TNF superfamily and is expressed on activated T lymphocytes, NK cells, monocytes, and granulocytes. CD178 is also expressed on parenchymal cells of the retina and cornea, retinal pigment epithelial cells, and testis. The extracellular region of FasL can be cleaved by matrix metalloproteinases (MMPs) to give rise to a 26 kD soluble protein. CD178 binds to CD95, a member of the TNFR superfamily, to induce apoptosis. CD95/CD95L interactions play an important role in the maintenance of peripheral tolerance and survival.
Other Names:
Fas Ligand (FasL), CD95L, TNFSF6
Structure:
TNF superfamily, type II glycoprotein, 38-42 kD, 26 kD soluble form
Distribution:
Activated T cells, NK cells, testis, eye, neutrophils, Clara type II cells
Function:
Apoptosis, immune privilege
Ligand Receptor:
CD95
Antigen References:
1. Suda, T., et al., 1997. J. Exp. Med. 12:204. 2. Kayagaki, N., et al., 1995. J. Exp. Med. 182:1777. 3. Tanaka, M., et al., 1995. EMBO J. 14:1129.
