The MG (Malachite Green) Phosphate Assay Kit is based on quantification of the green complex formed between Malachite Green, molybdate, and free orthophosphate. The rapid color formation from the reaction can be conveniently measured on a spectrophotometer (600-660 nm) or on a plate reader. The non-radioactive colorimetric assay kits have been optimized to offer superior sensitivity and prolonged shelf life. The assay is simple and fast, involving a single addition step for phosphate determination. Assays can be performed in tubes, cuvettes, or multi-well plates. The assays can be conveniently performed in 96 or 384-well plates for high-throughput screening of enzyme inhibitors. The kit has been used for assaying phosphatase, phospholipase or lipase, nucleoside triphosphatase (ATPase, GTPase, etc.) and phytase. The kit has the following components: • Highly purified Malachite Green reagent • Protein phosphatase inhibitor • Assay buffer • A 'mix and read' assay protocol that is compatible with HTS liquid handling instruments with Z' factors of 0.7 to 0.9.
Ref: Gurpal S. et al. (2003) Characterization of organic phosphorus in leachate from a grassland soil. Soil Biol.ogy & Biochemistry 35, 1317-1323; Cogan EB (1999) A robotics-based automated assay for inorganic and organic phosphates. Anal Biochem. 271, 29-35; Fisher DK and Higgins TJ (1994) A sensitive, high-volume, colorimetric assay for protein phosphatases. Pharm Res.11, 759-63; Henkel RD (1988) A microassay for ATPase. Anal Biochem. 169, 312-318.