Human TNF Receptor I protein, His tag

Tumor necrosis factor receptor superfamily member 1A (TNFRSF1A), also known as Tumor necrosis factor receptor 1 (TNFR1) and CD120a, is a ubiquitous membrane receptor that binds tumor necrosis factor-alpha (TNFα). This receptor can activate the transcription factor NF-κB, mediate apoptosis, and function as a regulator of inflammation. Antiapoptotic protein BCL2-associated athanogene 4 (BAG4/SODD) and adaptor proteins TRADD and TRAF2 have been shown to interact with this receptor, and thus play regulatory roles in the signal transduction mediated by the receptor. Besides, Granulin (GRN) has been identified as an interactor of TNFRSF1A, thus a binding ELISA assay was conducted to detect the interaction of recombinant human TNFRSF1A and recombinant human GRN. Briefly, TNFRSF1A were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to GRN-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-TNFRSF1A 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 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of of TNFRSF1A and GRN was in a dose dependent manner.