The ImmunoDiagnostics Rapid Human Cystatin C Immunoassay is a Solid Phase ELISA that can be run in an hour and detect Cystatin C in serum, plasma, or urine.
The ImmunoDiagnostics Rapid Human Cystatin C Kit (ELISA) is a reliable and reproducible tool for the quantitative detection of Cystatin C (also called Cystatin-3) production. The assay is intended to rapidly detect the target in subject serum, plasma, or urine.
Numerous application fields exist, but particularly:
- Nephrology: Cystatin C is effectively removed from urine by glomerular filtration. Therefore, its level in urine is anti-correlated with kidney function and research has suggested it to be a precise test for the glomerular filtration rate.
- Cardiovascular Disease: Several studies have found that increasing levels of Cystatin C are associated with an elevated risk multiple cardiovascular diseases.
- Oncology: In melanoma, a significant increase in Cystatin C was observed in subjects with metastatic disease and a smaller increase in with primary disease.
- Hepatology: Data suggest that levels of Cystatin C correspond to the progression of chronic liver disease. Further data show that it may be combined with albumin levels to study liver function in patients with cirrhosis
High quality, reproducible results in about an hour!
Cystatin C Antibody Coated Microstrips: One microplate, 12 strips with 8 wells each, 96 dry wells in total. The wells are coated with an anti- human Cystatin C antibody. The microplate is sealed in a foil bag. Any unused strips should be returned to the bag and resealed for future use.
Detection Antibody Solution: Concentrate (100x conc). One vial containing 0.12 mL of a horse radish peroxidase (HRP) conjugated polyclonal anti- human Cystatin C. Detection antibody should be diluted with only the 1X Assay Buffer needed.
Assay Buffer: Concentrate (5x conc). One vial containing 30 mL of buffer for dilution of the detection antibody. If precipitates are observed in the concentrated buffer, warm at 37oC until the precipitates disappear. Dilute 30 mL of 5x Assay Buffer with 120 mL diH2O to make a 1x concentration prior to use.
Human Cystatin C Standard: 15 ng of native human Cystatin C in a buffered protein base, lyophilized. Reconstitute with 1 mL 1x Assay Buffer to prepare 15 ng/mL standard stock. Prepare a serial dilution series with 1x Assay Buffer. The reconstituted standard stock can be aliquoted and stored at -20oC for one month. Avoid repeated freeze/thaw cycles.
Wash Buffer: Concentrate (10x conc). One vial containing 30 mL of 10x wash buffer. If precipitates are observed in the 10x solution, warm at 37oC until the precipitates disappear. Prepare 1x Wash Buffer my mixing 30 mL of 10x Wash Buffer with 270 mL diH2O.
Substrate Solution: One bottle containing 12 mL of TMB (3,3’,5,5’-Tetramethylbenzidine) substrate. Ready-to-use. Do not expose to light.
Stop Solution: One vial containing 12 mL of ready-to-use stop solution.
The Cystatin C ELISA is a quantitative sandwich enzyme immunoassay. Standard, samples, and any desired controls are pipetted into the wells pre-coated with solid-phase capture antibody. After washing away any unbound substances, an HRP-labelled detection antibody, specific for human Cystatin C, is added to the wells, and any unbound antibody is washed out. A TMB solution is added to develop color in proportion to the amount of human Cystatin C initially bound. The color development is stopped by addition of an acidifying solution and the optical density is determined. By plotting a standard curve versus measured absorbance, the amount of antigen in the sample can be calculated. The concentration of the antigen is expressed in ng/mL.
The kit contains calibrators and can be used in any laboratory equipped with a plate reader. The kit is optimized for the quantitative determination of human Cystatin C concentrations in serum, plasma, and urine samples. The test can provide results in about an hour.
Example only! Do not use for calculations
4 Parameter Curve
Log-log Regression Curve
Cystatin C, also called gamma trace, post-gamma-globulin, or neuroendocrine basic polypeptide, is primarily used in the determination of kidney function. Cystatin C is translated in all nucleated cells and is found in virtually all tissues and body fluids. Physiologically, it is a potent inhibitor of proteinases derived and stored in the lysozomes and, as such, it is a crucial inhibitor of extracellular cysteine proteases. Recent studies have indicated a potential role of cystatin c in predicting cardiovascular disease and it may play a role in amyloid-linked brain disorders (and is a potential target in Alzheimer’s Disease). Cystatin C has also been studied as a potential prognostic marker in several forms of cancer and corresponds to the progression of chronic liver disease and liver function in subjects with cirrhosis.
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