TNFRSF7 (Tumor necrosis factor receptor superfamily member 7), also known as CD27 antigen, is a member of the TNF-receptor superfamily. This receptor is thought to be involved in tumor necrosis factor-activated receptor activity by binding with TNFs. Thus, a binding ELISA assay was conducted to detect the ass°Ciation of TNFRSF7 with TNFa. Briefly, recombinant human TNFRSF7 were diluted serially in PBS with 0.01%BSA (pH 7.4). Duplicate samples of 100uL were then transferred to TNFa-coated microtiter wells and incubated for 2h at 37°C. Wells were washed with PBST and incubated for 1h with anti-TNFRSF7 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°C. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of TNFRSF7 with TNFa was shown in Figure 1 and this effect was in a dose dependent manner.