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Stock availability
In Stock
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Kit s description
Colorimetric measurement of human cystatin C levels
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Protein target
Cystatin C
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Scientific context
Cystatin C is a non-glycosylated protein of low molecular weight (13kDa) in the cystatin superfamily. Cystatin C is produced at a constant rate in all nucleated cells, secreted from cells and thus found in detectable amounts in most body fluids (1,2). Cystatin C belongs to the cysteine proteinase inhibitor group and is associated with several pathological conditions. Imbalance between Cystatin C and cysteine proteinases is associated with diseases such as inflammation, renal failure, cancer, Alzheimer’s disease, multiple sclerosis and hereditary Cystatin C amyloid angiopathy (1,3,4). Increased levels have been found in autoimmune diseases, with colorectal tumors and metastases, patients with inflammation and in patients on dialysis (5,6,7). Cystatin C is removed from blood plasma by glomerular filtration in the kidneys. It is reabsorbed by the proximal tubular cells and degraded. There is a linear relationship between the reciprocal Cystatin C concentration in plasma and the glomerular filtration rate (GFR). Cystatin C is suggested to be a better marker for GFR than the ubiquitous serum creatinine marker as its serum concentration is not affected by other factors such as age, gender and body mass and Cystatin C has higher sensitivity to detect a reduced GFR than creatinine determination (2,5). Low levels of Cystatin C are found with the breakdown of the elastic laminae and atherosclerosis and abdominal aortic aneurysm (8). There is evident association of Cystatin C levels with the incidence of myocardial infarction, coronary death and angina pectoris, presenting a risk factor for secondary cardiovascular events (9).
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Primary research area
Cell Signaling, Neuroscience, Neurodegeneration, Alzheimer's Disease, Cancer
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Category
Assay EIA Kits
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Other name
Cystatin 3 EIA Kit, gamma trace EIA Kit, post-gamma-globulin or neuroendocrine basic polypeptide EIA Kit
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Brand name
StressXpress®
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Detection system
Colorimetric Assay
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Assay format
Sandwich EIA (Enzyme Immunoassay)
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Main intended usage
EIA kit used to measure cystatin C levels in samples.
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Brief protocol
The Cystatin C EIA kit is designed to quantitatively measure human Cystatin C present in biological samples and tissue culture media. A human Cystatin C standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. Standards or diluted samples are pipetted into a clear microtiter plate coated with a monoclonal antibody to capture the Cystatin C present. After a 60 minute incubation, the plate is washed and a peroxidase conjugated Cystatin C monoclonal antibody is added. The plate is again incubated for 30 minutes and washed. Substrate is then added to the plate, which reacts with the bound Cystatin C Antibody Conjugate. After a third incubation, the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader capable of measuring 450 nm wavelength. The concentration of the Cystatin C in the sample is calculated, after making suitable correction for dilution, using software available with most plate readers.
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Kit contents
Clear 96 well Plate, Cystatin C Standard, StressXpress® Cystatin C Conjugate, Assay Buffer Concentrate, Wash Buffer Concentrate, TMB Substrate, Stop Solution, Plate Sealer
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Assay precision
Intra Assay Precision: Four human samples were diluted with Assay Buffer and run in replicates of 20 in an assay. The mean and precision of the calculated Cystatin C concentrations were: Sample 1- 7.72 ng/mL, 9.1% CV Sample 2- 5.08 ng/mL, 10.3% CV Sample 3- 4.42 ng/mL, 7.6% CV Sample 4- 0.88 ng/mL, 10.3% CV Inter Assay Precision: Four human samples were diluted with Assay Buffer and run in duplicates in ten assays run over multiple days by three operators. The mean and precision of the calculated Cystatin C concentrations were: Sample 1- 7.79 ng/mL, 8.4% CV Sample 2- 5.48 ng/mL, 10.2% CV Sample 3- 4.97 ng/mL, 11.1% CV Sample 4- 1.04 ng/mL, 12.40% CV
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Assay cross reactivity
Human
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Samples to be used with this kit
Serum, EDTA Plasma, Heparin Plasma, Urine, Tissue Culture Media
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Maximum samples to be used with this kit
40 samples in duplicate
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Assay duration
2 Hours
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Kit s sensitivity
0,058 ng/ml
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Assay detection limit
0,156 - 10 ng/ml
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Storage recommendations
4°C
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Shipping recommendations
Blue Ice
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Bibliography
1. Pergande M. and Jung K. Clin Chem. 1993 39(9):1885-90. 2. Newman D, et al. Kidney Int’l. 1995 Jan;47(1):312-8. 3. Levy E. Expert Rev. Neurotherapeutics 2008 May;8(5):687-9. 4. Nakashima I, et al. Ann Neurol. 2007 Aug;62(2):197-200. 5. Uchida K. and Gotoh, A. Clin Chim Acta. 2002 Sep;323:121-8. 6. Pucci L, et al. Clin Chem. 2007 Mar;53(3):480-8. 7. Stabuc B, et al. Clin Chem. 2000 Feb;46(2):193-7. 8. Shlipak M, et al. NEJM 2005 May;352(20):2049-60. 9. Koenig W, et al. Clin Chem. 2005 Feb;51(2):321-7.
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Release date
17-Sep-2014
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PubMed number
Refer to PubMed
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Tested applications
No data available / currently testing
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Tested reactivity
No data available / currently testing
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Resources available upon request
Kit Booklet, MSDS
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Product image link
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Product image legend
Typical Standard Curve for the Cystatin C EIA Kit (Enzyme Immunoassay) StressXpress® - SKT-219. Assay Type: Sandwich EIA. Detection Method: Colorimetric Assay. Assay Range: 0.156 - 10 ng/ml. | Serum linearity was determined by taking two human serum samples diluted 1:100, one with a low diluted Cystatin C level of 4.94 ng/mL and one with a higher diluted level of 12.31 ng/mL and mixing them in the ratios given below. Urine linearity samples were diluted 1:16. One sample with a value of 1.64 ng/mL was mixed in ratios with a sample of 3.72 ng/mL. The diluted serum and urine samples were mixed 4:1, 3:2, 2:3 and 1:4 and run in the assay. The measured concentrations were compared to the values previously determined. Graph of the Typical Standard Curve for the Cystatin C EIA Kit StressXpress - SKT-219 | Graph of the Linearity Recovery for the Cystatin C EIA Kit StressXpress - SKT-219
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Warnings
Non-hazardous
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Country of production
Canada
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Total weight kg
2
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Net weight g
300
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Description
This ELISA test kit or EIA Enzyme immuno assay is an enzyme linked immuno assay supplied as coated 96 well plates with antigen or antibody and needs to be stored at +4°C.
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Properties
MeSH E05 478 566 350 170 Enzyme-Linked Immunosorbent Assay
