Description
Principle of the Assay: Salusin Beta ELISA kit applies the competitive enzyme immunoassay technique utilizing a monoclonal anti-Salusin Beta antibody and an Salusin Beta-HRP conjugate. The assay sample and buffer are incubated together with Salusin Beta-HRP conjugate in pre-coated plate for one hour. After the incubation period, the wells are decanted and washed five times. The wells are then incubated with a substrate for HRP enzyme. The product of the enzyme-substrate reaction forms a blue colored complex. Finally, a stop solution is added to stop the reaction, which will then turn the solution yellow. The intensity of color is measured spectrophotometrically at 450nm in a microplate reader. The intensity of the color is inversely proportional to the Salusin Beta concentration since Salusin Beta from samples and Salusin Beta-HRP conjugate compete for the anti-Salusin Beta antibody binding site. Since the number of sites is limited, as more sites are occupied by Salusin Beta from the sample, fewer sites are left to bind Salusin Beta-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The Salusin Beta concentration in each sample is interpolated from this standard curve