The ImmunoDiagnostics CRP immunoassay is a Solid Phase ELISA designed to measure levels of CRP in human serum or plasma.
The ImmunoDiagnostics High Sensitive Human CRP kit (ELISA) is a reliable and reproducible tool for the quantitative determination of serum or plasma levels of human CRP.
Numerous application fields exist for CRP, but include:
- Hepatology: Research has shown that elevated CRP is a feature of non-alcoholic fatty liver disease (NAFLD) as well as nonalcoholic steohepatitis (NASH). Additionally, liver failure is one of the few known factors that can alter CRP production.
- Oncology: Elevated levels of CRP associated with chronic inflammation are associated with an elevated risk of developing cancer.
- Cardiovascular Disease and Metabolic Syndrome: Levels of CRP are associated with levels of risk of coronary heart disease when combined with other tests. Also, recent research indicates that higher levels of basal CRP lead to an increased risk for diabetes, hypertension, and cardiovascular diseases (CVD).
CRP Antibody Coated Microstrips: One microplate, 12 strips with 8 wells each, 96 dry wells in total. The wells are coated with mouse monoclonal anti-human CRP 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: One vial containing 0.12 mL of 100x Detection Antibody Concentrate. The antibody is a mouse monoclonal anti-human CRP conjugated to horseradish peroxidase. Dilution should be carried out by diluting with 1x Assay Buffer
Assay Buffer: One bottle containing 30 mL of 5x Assay Buffer Concentrate. If precipitates are observed in the concentrated buffer, heat at 37oC until the precipitate disappears. Dilute 30 mL of concentrated buffer with 120 mL of diH2O to make the 1x Assay Buffer prior to use.
Human CRP Standard: 0.3 mL of 100 ng/mL native human CRP in a buffered protein base solution.
Wash Buffer: 40 mL of 10x Wash Buffer Concentrate. If precipitates are observed in the concentrated buffer, heat at 37oC until the precipitate disappears. Dilute 40 mL concentrate with 360 mL diH2O.
Substrate Solution: One bottle of 12 mL of TMB (3,3’,5,5’-Tetramethybenzidine) substrate. Ready to Use. Do not expose to light
Stop Solution: One vial containing 12 mL of ready to use stop solution.
The hsCRP ELISA is a quantitative sandwich enzyme immunoassay. Standards, samples, and any desired controls are pipetted into the wells pre-coated with a solid phase capture antibody. After washing away any unbound substances, an HRP-labelled detection antibody, specific for human CRP, is added to the wells and subsequently washed out. A TMB substrate solution is added to develop color in proportion to the amount of human CRP initially bound to the well. The color development is stopped by acidification and the optical density is determined at 450 nm. By plotting a standard curve against corrected absorbance, the amount of antigen in the sample may 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 CRP concentration in serum and plasma in 2.5 hours. Typically, serum or plasma requires a 200 fold dilution (in 1x Assay Buffer) for this assay.
Log-log Regression Curve
C-Reactive Protein (CRP) is a circulating protein mainly secreted from the liver and synthesized in response to different factors released by macrophages and adipocytes. Its physiological role is to bind to lysophosphatidylcholine—found on the surface of dead cells, dying cells, and some bacteria—to trigger the complement system. While normally found in very low levels, its production is greatly increased in response to inflammation, infection, trauma, necrosis, and malignancy. An elevated basal level of CRP is associated with an increased risk of diabetes, hypertension, cardiovascular and coronary artery disease, and potentially some cancers.
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