Human RANK protein, His tag

RANK tumor necrosis factor receptor superfamily member 11A protein (TNFRSF11A) also known as receptor Activator of Nuclear Factor κ B (RANK) or TRANCE Receptoris a member of the tumor necrosis factor receptor (TNFR) molecular sub-family. TNFRSF11A is the receptor for RANK-Ligand (RANKL) and part of the RANK/RANKL/OPG signaling pathway that regulates osteoclast differentiation and activation. It is associated with bone remodeling and repair, immune cell function, lymph node development, thermal regulation, and mammary gland development. Besides, TNF Receptor Associated Factor 5 (TRAF5) has been identified as an interactor of TNFRSF11A, thus a binding ELISA assay was conducted to detect the interaction of recombinant human TNFRSF11A and recombinant human TRAF5. Briefly, TNFRSF11A were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to TRAF5-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-TNFRSF11A 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 TNFRSF11A and TRAF5 was in a dose dependent manner.