Active Death receptor 5 (DR5)

TNFRSF10B (Tumor necrosis factor receptor superfamily member 10B) is a member of the TNF-receptor superfamily, and contains an intracellular death domain. By binding to certain ligand, this receptor transduces an apoptosis signal. A binding ELISA assay was conducted to detect the association of FAS with TNFa. Briefly, FAS were diluted serially in PBS, with 0.01%BSA (pH 7.4). Duplicate samples of 100uL FAS were then transferred to TNFa-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-FAS pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 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 TNFa was shown in Figure 1, and this effect was in a dose dependent manner.