Protein phosphatases have received great attention as drug-screening targets. FDP is a highly sensitive fluorogenic substrate for most phosphatases, i.e., alkaline phosphatases, protein tyrosine phosphatases and serine/threonine phosphatases. The phosphatase-induced hydrolysis of FDP yields an intense fluorescent product that has excitation wavelength around 490 nm, and emission maximum around 520 nm. The fluorescence of FDP hydrolyzed product is strengthened when pH > 7.0. For some protein phosphatase requiring slightly acidic buffer for reaction, a pH-adjusting stop solution should be added before reading the fluorescence signal. The kit can be used for characterizing kinetics of enzyme reaction and high throughput screening of protein phosphatase inducers and inhibitors.
Component A: FDP, Ex/Em= 485±20/528±20 nm upon phosphate group removal: 1 vial
Component B: Assay buffer, pH 6.5: 60 ml
Component C: 10X Lysis buffer: 50 ml
Component D: Triton X-100: 500 µL
Component E: Stop solution: 30 ml
Component F: 1 M DTT: 100 µl
Component G: DMSO: 500 µl
Storage & stability
Store components A and F at -20°C. Component B, C, D, E and G can be stored at room temperature for convenience.