Description
Principle of the assay: human MICA ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for MICA has been precoated onto 96-well plates. Standards(NSO, E24-W307) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for MICA 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 MICA amount of sample captured in plate.
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