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EK000078-EK0812: Human MICA ELISA Kit
Range 62.5pg/ml-4000pg/ml
Sensitivity < 10pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitative detection of human MICA in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
MHC class I polypeptide-related sequence A is a protein that in humans is encoded by the MICA gene.1 The MICA gene encodes a 383-amino acid polypeptide with a predicted mass of 43 kD.The MICA and MICB genes occur in a 200-kb region spanning the TNFA and TNFB cluster at 6p21.3.2 MICA and the closely related MICB were recognized by intestinal epithelial T cells expressing diverse V-delta-1 gamma/delta TCRs.3 The MICA protein product is expressed on the cell surface, although unlike canonical class I molecules does not seem to associate with beta-2-microglobulin. It is further distinguished by its unusual exon-intron organization and preferential expression in fibroblasts and epithelial cells. It is thought that MICA functions as a stress-induced antigen that is broadly recognized by NK cells, NKT cells, and most of the subtypes of T cells. MICA and other members of this family may have been selected for specialized functions that are either ancient or derived from those of typical MHC class I genes, in analogy to some of the nonclassic mouse H-2 genes. The standard product used in this kit is recombinant MICA, which is composed of 286 amino acids with the molecular mass of 59KDa.
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. Bahram S, Bresnahan M, Geraghty DE, Spies T (Aug 1994). "A second lineage of mammalian major histocompatibility complex class I genes". Proc Natl Acad Sci U S A 91 (14): 6259-63.
2. Nalabolu, S. R.; Shukla, H.; Nallur, G.; Parimoo, S.; Weissman, S. M. : Genes in a 220-kb region spanning the TNF cluster in human MHC. Genomics 31: 215-222, 1996.
3. Groh, V.; Steinle, A.; Bauer, S.; Spies, T. : Recognition of stress-induced MHC molecules by intestinal epithelial gamma-delta T cells. Science 279: 1737-1740, 1998.
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