Fas ligand (FasL or CD95L) is a type-II transmembrane protein that belongs to the tumor necrosis factor (TNF) family. Its binding with its receptor induces apoptosis. The human FASL gene consists of approximately 8kb and is split into 4 exons. Fas ligand/receptor interactions play an important role in the regulation of the immune system and the progression of cancer. Fas ligand or FasL is a homotrimeric type II transmembrane protein. It signals through trimerization of FasR, which spans the membrane of the target cell. This trimerization usually leads to apoptosis, or cell death. Soluble Fas ligand is generated by cleaving membrane-bound FasL at a conserved cleavage site by the external matrix metalloproteinase MMP-7. Intended Use: For quantitative detection of human FASL in cell culture supernates, serum and plasma (heparin, EDTA, citrate). Range: 15.6-1000pg/ml Sensitivity: < 2pg/ml Specificity: Natural and recombinant human FASL Crossreactivity: No detectable crossreactivity with other relevant proteins Kit Components: 143829A: Microtiter Strip, 1x96 wells 143829B: FASL Standard, 2x10ng 143829C: Anti-human FASL (Biotin), 130ul USB Cat No Kit Component 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (1:100 dilution) 143697: Sample diluent buffer, 1x30ml 143698: Antibody diluent buffer, 1x12ml 143699: ABC diluent buffer, 1x12ml 143700: TMB color developing agent, 1x10ml 143701: TMB stop solution, 2N H2SO4, 1x10ml Storage and Stability: Store *143829B powder at 4C. Once reconstituted store at 4C for up to 12 hours or at -20C for up to 48 hours. Store other components at 4C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Assay Summary: 1. Add 100ul of samples and standards and incubate the plate at 37C for 90 min. Do not wash. 2. Add 100ul biotinylated antibody and incubate the plate at 37C for 60 min. Wash plate 3 times with 0.01M TBS or PBS. 3. Add ABC working solution and incubate the plate at 37C for 30min. Wash plate 5 times with 0.01M TBS or PBS. 4. Add TMB color developing agent and incubate the plate at 37C in dark for 20-25 min. 5. Add 100ul TMB stop solution and read.
