AffiELISA® Rat Phospho-Akt Kit: High Sensitivity S473 ELISA Detection

The AffiELISA® Rat Phospho-Akt Kit uses Sandwich enzyme immunoassay. The provided microtiter plate comes pre-coated with an antibody specific to Phospho-Akt (S473) . To perform the assay, standards or samples are added to designated wells on the microtiter plate. Subsequently, a biotin-conjugated antibody specific to Phospho-Akt (S473) is introduced. Avidin conjugated to Horseradish Peroxidase (HRP) is then added to each well, followed by an incubation period. After TMB substrate solution is added, only those wells that contain Phospho-Akt (S473) , biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. By comparing the optical density (OD) of the samples to the standard curve, the concentration of Phospho-Akt (S473) in the samples can be determined.