

- CAN:
- EU:
- USA: For Research Use Only. Not For Use In Diagnostic Procedures.
The hs-CRP ELISA is a two-step capture or ‘sandwich’ type immunoassay. The assay makes use of two highly specific monoclonal antibodies: A monoclonal antibody specific for CRP is immobilized onto the microplate and another monoclonal antibody specific for a different epitope of CRP is conjugated to horse radish peroxidase (HRP conjugate). In the first incubation step, CRP present in the specimen samples, calibrators and controls is bound by the antibody immobilized onto the microplate. Excess and unbound materials are removed by a washing step.
In the second incubation step, HRP conjugate antibody (HRP conjugate) is added, which binds specifically to any immobilized CRP, thus forming a sandwich complex. Unbound HRP conjugate is removed by a washing step. Next, the TMB substrate (enzyme substrate) is added which reacts with HRP to form a blue coloured product that is directly proportional to the amount of CRP present. The enzymatic reaction is terminated by the addition of the stopping solution, converting the colour from blue to yellow. The absorbance is measured on a microplate reader at 450 nm. A set of calibrators is used to plot a calibrator curve from which the amount of CRP in specimen samples and controls can be directly read.