DuoSet IC Phosphatase Activity Assay Principle

Microplates are coated with the capture antibody provided in the assay. Samples containing phosphatase are added.

Any phosphatase present is bound by the immobilized antibody. Unbound materials are washed away.

A synthetic phosphopeptide substrate is added that is dephosphorylated by active phosphatase to generate free phosphate and unphosphorylated peptide. The free phosphate is detected by a sensitive dye binding assay using malachite green. By calculating the rate of phosphate release, the activity of the specific phosphatase of interest is determined.