Description
Principle of the Assay: This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-FAS polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-FAS polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the FAS amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of FAS can be calculated.
Background: Fas, also known as CD95, APO-1 and TNFRSF6, is a member of the TNF-receptor superfamily. This receptor contains a death domain. It has been shown to play a central role in the physiological regulation of programmed cell death, and it has been implicated in the pathogenesis of various malignancies and diseases of the immune system. The interaction of Fas with its ligand allows the formation of a death-inducing signaling complex that includes Fas-associated death domain protein (FADD), caspase 8, and caspase 10. The autoproteolytic processing of the caspases in the complex triggers a downstream caspase cascade and leads to apoptosis. This receptor has been also shown to activate NF-kappaB, MAPK3/ERK1, and MAPK8/JNK, and it is found to be involved in transducing the proliferating signals in normal diploid fibroblast and T cells