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EK000046-EK0530: Human Soluble TNFRII ELISA Kit
Range 7.8pg/ml-500pg/ml
Sensitivity < 2 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitative detection of human sTNFRII in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Tumor necrosis factor receptor p75 (TNF-R p75, or TNF-R2) is a 75-kDa type I transmembrane protein expressed predominantly on cells of hematopoietic lineage. TNF-R p75 belongs to the TNF receptor superfamily characterized by cysteine-rich extracellular regions composed of three to six disulfide-linked domains.1 The tumor necrosis factor receptor ? (TNFRII gene localizes to 1p36. 2, a genomic region characteristically deleted in neuroblastomas and other malignancies. In addition, TNFRII is the principal mediator of the effects of TNF on cellular immunity, and it may cooperate with TNFRII in the killing of nonlymphoid cells.2 The standard product used in this kit is recombinant human sTNFR, consisting of 24-206 amino acid sequence with the molecular mass of 20KDa.
Principle
The ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. The target specific antibodies are precoated onto 96-well plates. The target from the sample is bound to the microwell. The biotinylated target specific detection antibodies are added to the microwells and followed by washing with the PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with the PBS or TBS buffer. TMB, the HRP (horseradish peroxidase) substrate, is used to visualize color change resulting from the HRP enzymatic reaction. TMB is catalyzed by HRP to produce the blue color. The color changes into yellow after the acidic stop solution is added. The density of the yellow color is proportional to the target amount from the sample captured in the microwells.
Reference
1. Santee, S. M.; Owen-Schaub, L. B. Human tumor necrosis factor receptor p75/80 (CD120b) gene structure and promoter characterization. J. Biol. Chem. 271: 21151-21159, 1996.
2. Beltinger, C. P.; White, P. S.; Maris, J. M.; Sulman, E. P.; Jensen, S. J.; LePaslier, D.; Stallard, B. J.; Goeddel, D. V.; de Sauvage, F. J.; Brodeur, G. M. Physical mapping and genomic structure of the human TNFR2 gene. Genomics 35: 94-100, 1996.
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