Description
Principle of the assay: human TNFsRIELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for TNFsRI has been precoated onto 96-well plates. Standards and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for TNFsRI is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human TNFsRI amount of sample captured in plate.
Background: TNFsRI(also known as Solube tumor necrosis factor receptor 1) is a cytokine receptor that binds tumor necrosis factors (TNF). In contrast, the p60 (TNFRSF1A) and p80 (TNFRSF1B) TNFA receptors self-assemble through a distinct functional domain in the TNFR extracellular domain, termed the pre-ligand assembly domain (PLAD), in the absence of ligand. Deletion of the PLAD results in monomeric presentation of p60 or p80. Flow cytometric analysis showed that efficient TNFA binding depends on receptor self-assembly. They also found that other members of the TNF receptor superfamily, including the extracellular domains of TRAIL (TNFRSF10A), CD40, and FAS (TNFRSF6), all self-associate but do not interact with heterologous receptors. By Southern blot analysis of human/Chinese hamster somatic cell hybrid DNA, the TNFR1 gene was mapped to 12pter-cen. And by nonradioactive in situ hybridization that the type 1 receptor (the p55 TNF receptor) is encoded by a gene located on chromosome 12p13.2