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EK000136-EK0528: Human TNFs RI ELISA Kit
Range 15.6pg/ml-1000pg/ml (Body fluids, tissue lysates or cell culture supernates) 7.8pg/ml-500pg/ml (Human sera)
Sensitivity < 1 pg/ml
Specificity: No detectable cross-reactivity with any other cytokine.
Application: For quantitative detection of human TNFs RI in sera, body fluids, tissue lysates or cell culture supernates.
Expiration: Four months at 4°C and eight months at -20°C.
Background
TNFs RI (also known as Solube tumor necrosis factor receptor 1) is a cytokine receptor that binds tumor necrosis factors (TNF). Chan et al. (2000) found that, in contrast, the p60 (TNFRSF1A) and p80 (TNFRSF1B; 191191) 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; 603611), CD40 (109535), and FAS (TNFRSF6; 134637), all self-associate but do not interact with heterologous receptors. By Southern blot analysis of human/Chinese hamster somatic cell hybrid DNA, Milatovich et al. (1991, 1991) mapped the TNFR1 gene to 12pter-cen. Derre et al. (1991) found by nonradioactive in situ hybridization that the type 1 receptor (the p55 TNF receptor) is encoded by a gene located on chromosome 12p13.2.
Principle
TNFs RI ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human TNFs RI specific-specific monoclonal antibodies (clone No. 28401.111) were precoated onto 96-well plates. The human specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells 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 TNFs RI amount of sample captured in plate.
Reference
1 Chan, F. K.-M., Chun, H. J., Zheng, L., Siegel, R. M., Bui, K. L., Lenardo, M. J. A domain in TNF receptors that mediates ligand-independent receptor assembly and signaling. Science 288: 2351-2354, 2000.
2. Milatovich, A., Song, K., Heller, R. A., Francke, U. The genes for two tumor necrosis factor receptors, TNFR1 and TNFR2, map to human chromosomes 12 (12pter-cen) and 1 (1pter-p32). Cytogenet. Cell Genet. 58: 1980-only, 1991.
3. Derre, J., Kemper, O., Cherif, D., Nophar, Y., Berger, R., Wallach, D. The gene for the type 1 tumor necrosis factor receptor (TNF-R1) is localized on band 12p13. Hum. Genet. 87: 231-233, 1991.
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