Active Factor Related Apoptosis (FAS)

FAS (Tumor necrosis factor receptor superfamily member 6) belongs to the tumor necrosis factor receptor superfamily. FAS contains a death domain, which has been shown to play a central role in the physiological regulation of programmed cell death. A binding ELISA assay was conducted to detect the association of FAS with FASL. Briefly, FASL were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 ul FASL were then transferred to FAS-coated microtiter wells (1ug/ml, 100ul/well) and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-FASL pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of FAS and FASL was shown in Figure 1, and this effect was in a dose dependent manner, the EC50 was approximately 0.012 ug/mL.