Principle of Assay
Bovine alpha Lactalbumin ELISA Kit (A87162) is a Competitive ELISA for the quantitative measurement of bovine alpha Lactalbumin in serum, plasma, tissue homogenates, and other biological fluids. An antibody specific for alpha Lactalbumin has been pre-coated onto a 96-well microtiter plate. During the incubation, alpha Lactalbumin in the sample or standard competes with a fixed amount of Biotinylated-alpha Lactalbumin. Following incubation, Biotinylated-alpha Lactalbumin and free alpha Lactalbumin 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 alpha Lactalbumin 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 alpha Lactalbumin in the sample.
