Description
Principle of the assay: mouse Cystatin C ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for Cystatin C has been precoated onto 96-well plates. Standards(NSO, M1-A140) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Cystatin C 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 mouse Cystatin C amount of sample captured in plate.
Background: Cystatin C or cystatin 3 (formerly gamma trace, post-gamma-globulin or neuroendocrine basic polypeptide), a protein encoded by the CST3 gene, was originally described as a constituent of normal cerebrospinal fluid (CSF) and of urine from patients with renal failure. Cystatin 3 has a low molecular weight (approximately 13.3 kilodaltons), and it is removed from the bloodstream by glomerular filtration in the kidneys. In mouses, all cells with a nucleus (cell core containing the DNA) produce cystatin C as a chain of 120 amino acids. It is found in virtually all tissues and bodily fluids. Cystatin C, which belongs to the type II cystatin gene family, is a potent inhibitor of lysosomal proteinases (enzymes from a special subunit of the cell that break down proteins) and probably one of the most important extracellular inhibitors of cysteine proteases (it prevents the breakdown of proteins outside the cell by a specific type of protein degrading enzymes). Moreover, cystatin C is involved in network reorganization in the epileptic dentate gyrus