Description
Principle of the Assay: The ELISA is based on the competitive binding enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with an antibody specific to PEPCK-M, During the reaction, PEPCK-M in the sample or standard competes with a fixed amount of biotin-labeled PEPCK-M for sites on a pre-coated Monoclonal antibody specific to PEPCK-M. Excess conjugate and unbound sample or standard are washed from the plate. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm +/- 2 nm. The concentration of PEPCK-M in the samples is then determined by comparing the O.D. of the samples to the standard curve