Description
The Eagle Biosciences hsCRP ELISA Assay Kit is based on the principle of a solid phase enzyme-linked immunosorbent assay.24 The assay system utilizes a unique monoclonal antibody directed against a distinct Antigensic 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 enzyme-linked 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 450 nm