High Sensitivity Enzyme Immunoassay for the Quantitative Determination of C-Reactive Protein Concentration in Human Serum

for in vitro diagnostic use

Product Description
C- Reactive protein (CRP) was identified by Tilet and Francis (1930) in the plasma of patients with pneumonia, and was named for its ability to bind and precipitate the C-polysaccharide of pneumococcus. It is an alpha globulin with a molecular mass of approximately 110,000 to 140,000 daltons, and is composed of five identical subunits, which are noncovalently assembled as a cyclic pentamer. CRP is synthesized in the liver and is normally present as a trace constituent of serum or plasma at levels less than 0.3mg/dl. Its physiological roles are numerous and varied, but with several functions similar to those of immunoglobulins, CRP appears to function in host defense.

Principle
The hsCRP ELISA is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay system utilizes a unique monoclonal antibody directed against a distinct antigenic determinant on the on the CRP molecule. This mouse monoclonal anti-CRP antibody is used for solid phase immobilization (on the microtiter wells).
A goat anti-CRP antibody is in the antibody-enzyme (horseradish peroxidase) conjugate solution. The test sample is allowed to react simultaneously with the two antibodies, resulting in the CRP molecules being sandwiched between the solid phase and enzymelinked antibodies.
After a 45-minute incubation at room temperature, the wells are washed with water to remove unbound labeled antibodies.
A tetramethylbenzidine (TMB) reagent is added and incubated for 20 minutes, resulting in the development of blue color. The color development is stopped with the addition of 1N HCl changing the color to yellow. The concentration of CRP is directly proportional the color intensity of the test sample.
Absorbance is measured spectrophotometrically at 450nm.

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