Principle of Assay
Human Salivary alpha Amylase ELISA Kit (A77676) is a Competitive ELISA for the quantitative measurement of human Salivary alpha Amylase in serum, plasma, tissue homogenates, and other biological fluids. An antibody specific for Salivary alpha Amylase has been pre-coated onto a 96-well microtiter plate. During the incubation, Salivary alpha Amylase in the sample or standard competes with a fixed amount of Biotinylated-Salivary alpha Amylase. Following incubation, Biotinylated-Salivary alpha Amylase and free Salivary alpha Amylase in the sample or standard are washed from the plate. HRP-Streptavidin (SABC) is then added to each well and incubated. Unbound SABC is washed away with wash buffer. TMB substrate solution is added and after incubation the enzyme-substrate reaction is terminated by the addition of sulfuric acid stop solution. The colour change is measured spectrophotometrically at a wavelength of 450nm and the concentration of Salivary alpha Amylase in the samples is then determined by comparing the O.D. 450nm of the samples to the standard curve. O.D. 450nm is inversely proportional to the concentration of Salivary alpha Amylase in the sample.
