ProFluor Ser/Thr PPase Assay, 4 plate Item ID: PMV1260

fluorescence intensity,rhodamine 110,phosphatase assays,calcineurin,PP1,PP2A,PP2B,PP2C,cdc25,profluor,pro fluor,pro-fluor,v126

The ProFluor Ser/Thr PPase Assay measures purified serine/threonine protein phosphatase activity in a multiwell plate format and is ideal for high-throughput applications.

The ProFluor Ser/Thr PPase Assay measures purified serine/threonine protein phosphatase activity in a multiwell plate format and involves add-mix-read steps only---ideal for high-throughput applications. The assay works with protein phosphatase 1 (PP1), PP2A, PP2B and PP2C. The assay begins with a standard phosphatase reaction performed with a provided phosphorylated bisamide rhodamine 110 peptide substrate (S/T PPase R110 Substrate) and Control AMC Substrate that serves as a control for compounds that may inhibit the protease reaction. Following the phosphatase reaction, a termination buffer containing a protease reagent is added, which simultaneously stops the phosphatase reaction and removes amino acids specifically from the nonphosphorylated substrate, liberating highly fluorescent rhodamine 110. Phosphorylated substrate, however, is resistant to digestion by the protease reagent and remains nonfluorescent. Thus, fluorescence intensity measured in this assay is directly correlated with phosphatase activity. The assay produces excellent Z' values (>0.8) in either 96- or 384-well plate formats and easily distinguishes known phosphatase inhibitors from other compounds.