Description
Principle of the Assay: PYD ELISAkit applies the quantitative sandwich enzyme immune-assay technique. The microtiter plate has been pre-coated with a monoclonal antibody specific for PYD. Standards or samples are then added to the microtiter plate wells and PYD if present, will bind to the antibody pre-coated wells. In order to quantitatively determine the amount of PYD present in the sample, a standardized preparation of horseradish peroxidase (HRP) -conjugated polyclonal antibody specific for PYD are added to each well to ''sandwich'1 the PYD 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 PYD 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 spectrophotomehically at a wavelength of 450 nm. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The PYD concentration in each sample is interpolated from this standard curve