Description
This Human Cystatin C ELISA Kit is intended for quantitative detection of Human Cystatin C in cell culture supernates, serum, plasma (heparin, EDTA), saliva, urine and human milk. Strip well format. Reagents for up to 96 tests.
This human Cystatin C ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Cystatin C has been precoated onto 96-well plates. Standards (NSO, M1-A146) 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 human Cystatin C amount of sample captured in plate.
The capture antibody is a monoclonal antibody from mouse, the detection antibody is a biotinylated polyclonal antibody from goat. Expression system for standard: 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.1 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 humans, 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 proteinases2 (enzymes from a special subunit of the cell that break down proteins) and probably one of the most important extracellular inhibitors of cysteine proteases3 (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.4