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The PKA kinase activity kit allows relative quantification of PKA kinase activity in different sample types (purified, partially purified and crude enzyme preparations) from any species. A peptide substrate is coated on the microtiter plate and in the presence of active PKA enzyme and ATP it becomes phosphorylated. Incubation with a phosphospecific antibody followed by a peroxidase conjugated secondary antibody and a TMB substrate allows detection and quantification of the phosphorylated peptide. The generated color intensity is measured in a microplate reader at 450nm and it is proportional to PKA phosphotransferase activity.