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EK000092-EK0679: Mouse Cystatin C ELISA Kit
Range 0.312ng/ml-20ng/ml
Sensitivity < 10 pg/ml
Specificity: No detectable cross-reactivity with any other cytokine.
Application: For quantitative detection of mouse Cystatin C in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: Four months at 4°C and eight months at -20°C.
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.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 The standard product used in this kit is recombinant human Cystatin C, consisting of 120 amino acids with the molecular mass of 15KDa.
Principle
This assay is a sandwich ELISA designed for the quantitative detection of human cystatin C in samples in 1 hour. The immunoplate is pre-coated with antibody specific to human cystatin C. Standards and samples are pipetted into the wells and any human cystatin C present is sandwiched by the immobilised antibody and a second horseradish peroxidase (HRP)-linked antibody specific to human cystatin C that is co-incubated with the samples. After wash step to remove any unbound substances, the HRP substrate solution is added and colour develops in proportion to the amount of human cystatin C bound initially. The assay is stopped and the optical density of the wells determined using a micro-plate reader. Since the increases in absorbance are directly proportional to the amount of captured human cystatin C, the unknown sample concentration can be interpolated from a reference curve included in each assay.
Reference
1. Grubb, A.; Lofberg, H. : Human gamma-trace, a basic microprotein: amino acid sequence and presence in the adenohypophysis. Proc. Nat. Acad. Sci. 79: 3024-3027, 1982.
2. Pirttila, T. J.; Manninen, A.; Jutila, L.; Nissinen, J.; Kalviainen, R.; Vapalahti, M.; Immonen, A.; Paljarvi, L.; Karkola, K.; Alafuzoff, I.; Mervaala, E.; Pitkanen, A. : Cystatin C expression is associated with granule cell dispersion in epilepsy. Ann. Neurol. 58: 211-223, 2005.
3. Abrahamson, M. : Human cysteine proteinase inhibitors: isolation, physiological importance, inhibitory mechanism, gene structure and relation to hereditary cerebral hemorrhage. Scand. J. Clin. Lab. Invest. 48 (suppl. 191): 21-31, 1988.
4. Pirttila, T. J.; Manninen, A.; Jutila, L.; Nissinen, J.; Kalviainen, R.; Vapalahti, M.; Immonen, A.; Paljarvi, L.; Karkola, K.; Alafuzoff, I.; Mervaala, E.; Pitkanen, A. : Cystatin C expression is associated with granule cell dispersion in epilepsy. Ann. Neurol. 58: 211-223, 2005.
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