Description
Intended Uses: This ASMA ELISA kit is a 1.5 hour solid-phase ELISA designed for the quantitative determination of Rabbit ASMA.
Principle of the Assay: ASMA ELISA kit applies the competitive enzyme immunoassay technique utilizing Smooth Muscle antigen and an ASMA-HRP conjugate. The assay sample and buffer are incubated together with ASMA-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 ASMA concentration since ASMA from samples and ASMA-HRP conjugate compete for the Smooth Muscle antigen binding site. Since the number of sites is limited, as more sites are occupied by ASMA from the sample, fewer sites are left to bind ASMA-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The ASMA concentration in each sample is interpolated from this standard curve