Description
Principle of the Assay: CR ELISA kit applies the quantitative sandwich enzyme imniunoassay technique. The microtiter plate has been pre-coated with a monoclonal antibody specific for CR. Standards or samples are then added to the microtiter plate wells and CR if present, will bind to the antibody pre-coated wells. In order to quantitatively determine the amount of CR present in the sample, a standardized preparation of horseradish peroxidase (HRP) -conjugated polyclonal antibody specific for CR are added to each well to "sandwich" the CR immobilized on the plate. The microtiter plate undergoes incubation, and then the wells are thoroughly washed to remove all unbound components. Next, substrate solutions are added to each well. The enzyme (HRP) and substrate are allowed to react over a short incubation period. Only those wells that contain CR and enzyme-conjugated antibody will exhibit a change in color. The enzyme-substrate reaction is terminated by addition of a sulphuric acid solution and the color change is measured spectrophotonietrically at a wavelength of 450 mn. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The CR concentration in each sample is interpolated from this standard curve