Human TNF Receptor II protein, His tag (active)

GeneTex

Human TNF Receptor II protein (active)

9

Tumor necrosis factor receptor superfamily member 1B (TNFRSF1B), also known as tumor necrosis factor receptor 2 (TNFR2) and CD120b, is a membrane receptor that binds tumor necrosis factor-alpha (TNFalpha). This protein and TNF-receptor 1 form a heterocomplex that mediates the recruitment of two anti-apoptotic proteins, c-IAP1 and c-IAP2, which possess E3 ubiquitin ligase activity. Besides, Tumor Necrosis Factor Alpha (TNFa) has been identified as an interactor of TNFRSF1B, thus a binding ELISA assay was conducted to detect the interaction of recombinant human TNFRSF1B and recombinant human TNFa. Briefly, TNFRSF1B were diluted serially in PBS, with 0. 01% BSA (pH 7. 4). Duplicate samples of 100 microl were then transferred to TNFa-coated microtiter wells and incubated for 2h at 37C. Wells were washed with PBST and incubated for 1h with anti-TNFRSF1B 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 37C. Finally, add 50 microl stop solution to the wells and read at 450nm immediately. The binding activity of TNFRSF1B and TNFa was in a dose dependent manner. To tested cell apoptosis, A549 cells were seeded into triplicate wells of 96-well plates at a density of 2000 cells/well. and allowed to attach, replaced with serum-free overnight, then the medium was replaced with 2% serum standard DMEM including 1microg/ml TNFalpha and various concentrations of recombinant human TNFRSF1B. After incubated for 96h, cells were observed by inverted microscope and cell proliferation was measured by Cell Counting Kit-8 (CCK-8). Briefly, 10 microl of CCK-8 solution was added to each well of the plate, then the absorbance at 450nm was measured using a microplate reader after incubating the plate at 37C for 1-4 hours. Apoptosis of A549 cells had been inhibit after incubation with TNFRSF1B for 96h observed by inverted microscope . Cell viability was assessed by CCK-8 (Cell Counting Kit-8) assay after incubation with recombinant TNFRSF1B for 96h. And TNFRSF1B significantly suppress cell apoptosis induced by TNFalpha.

Functional Assay

Research Use Only

Unconjugated

Tumor necrosis factor receptor superfamily member 1B

The protein encoded by this gene is a member of the TNF-receptor superfamily. This protein and TNF-receptor 1 form a heterocomplex that mediates the recruitment of two anti-apoptotic proteins, c-IAP1 and c-IAP2, which possess E3 ubiquitin ligase activity. The function of IAPs in TNF-receptor signalling is unknown, however, c-IAP1 is thought to potentiate TNF-induced apoptosis by the ubiquitination and degradation of TNF-receptor-associated factor 2, which mediates anti-apoptotic signals. Knockout studies in mice also suggest a role of this protein in protecting neurons from apoptosis by stimulating antioxidative pathways. [provided by RefSeq, Jul 2008]

-20°C or -80°C,2°C to 8°C

12352202