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Human TNF Receptor II protein (active)

Human TNF Receptor II protein (active). GTX00235-pro
Human TNF Receptor II protein (active). GTX00235-pro
Human TNF Receptor II protein (active). GTX00235-pro
Human TNF Receptor II protein (active). GTX00235-pro

Cat No. GTX00235-pro

Application Functional Assay
Species Human
Package
10 μg ($259)
APPLICATION

Application Note

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 (TNFα). 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 μl were then transferred to TNFa-coated microtiter wells and incubated for 2h at 37ºC. 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 37ºC. Finally, add 50 μl 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 1μg/ml TNFα 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 μl 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 37ºC 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 TNFα.

Observed MW

50 kDa.
PROPERTIES

Form

Lyophilized powder

Buffer

Lyophilized from 20 mM Tris (pH 8.0) with 150 mM NaCl, 1 mM EDTA, 1 mM DTT, 0.01% SKL, 5% Trehalose, Proclin300. Reconstitute with 20 mM Tris and 150 mM NaCl (pH 8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

Storage

For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution.Avoid freeze-thaw cycles.

Region/Sequence

N-terminal His-Tag; Ala298~Leu441 (NP_001057.1)

Expression System

E. coli

Purity

> 95%

Endotoxin

< 1 EU/μg

Conjugation

Unconjugated

Note

For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
TARGET

Synonyms

TNF receptor superfamily member 1B,CD120b,TBPII,TNF-R-II,TNF-R75,TNFBR,TNFR1B,TNFR2,TNFR80,p75,p75TNFR

Background

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]

Database

Research Area

DATA IMAGES
Human TNF Receptor II protein (active). GTX00235-pro

GTX00235-pro Functional Assay Image

The suppression effect of GTX00235-pro Human TNF Receptor II protein (active) on TNF alpha induced apoptosis.
(A) A549 cells cultured in DMEM only contain 1 μg/ml TNF alpha for 96hrs.
(B) A549 cells cultured in DMEM contain 1 μg/ml TNF alpha and 1 ng/ml TNFRSF1B for 96hrs.

Human TNF Receptor II protein (active). GTX00235-pro

GTX00235-pro Functional Assay Image

Functional ELISA analysis of GTX00235-pro Human TNF Receptor II protein (active) which can bind immobilized TNFa protein.

Human TNF Receptor II protein (active). GTX00235-pro

GTX00235-pro Image

WB analysis of GTX00235-pro Human TNF Receptor II protein (active).

Human TNF Receptor II protein (active). GTX00235-pro

GTX00235-pro Image

SDS-PAGE analysis of GTX00235-pro Human TNF Receptor II protein (active).

REFERENCE
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REVIEW
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