Malachite Green Assay Lab Report

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Malachite Green Assay: The reaction that AGPase catalyzes produces PPi. The resulting PPi that is produced is hydrolyzed by an inorganic pyrophosphatase (Sigma Aldrich) to generate phosphate (Pi). Each reaction for the assay was conducted on a 96 well plate and the reaction mixture was 50µL and contained Pre-Mix which contained 50mM HEPPS, pH=8.0 (MP Biomedicals), 10mM MgCl2, 1.5mM FBP (Sigma Aldrich), and 0.2mg/mL Bovine Serum Albumin (Sigma Aldrich), and 1.5mU/µL yeast inorganic pyrophosphatase. ATP was added, whose final concentration was 1.5mM and the rest of the volume was the desired dilution of the protein solution in question, water, or pyrophosphate (Sigma Aldrich). Assays were initiated with ATP or water and incubated for 10 minutes

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