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A small aliquot of the assay was transferred to the solution of the assemblies at set time intervals. The fluorescence of the solution is measured after 20 min of incubation time. The initial concentration of sodium PPi in the assay was 1.6 mM [1.0 mM, One unit of pyrophosphatase will liberate 1.0 μmol of inorganic Pi per minute at pH 9.0 at 50°C The fluorescence recovery from the solution gradually decreased with increasing incubation time. Beyond a specific incubation time of the assay 3 min for EA200; 5 min for EA100; 1.8 min for EA70, 30 min EA 40, the fluorescence intensity of the solution remained constant, indicating that all of the PPi had been hydrolyzed by the enzyme. No further fluorescence recovery was observed
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A small aliquot of the assay was transferred to the solution of the assemblies at set time intervals. The fluorescence of the solution is measured after 20 min of incubation time. The initial concentration of sodium PPi in the assay was 1.6 mM [1.0 mM]. One "unit" of pyrophosphatase will liberate 1.0 μmol of inorganic Pi per minute at pH 9.0 at 50°C The fluorescence recovery from the solution gradually decreased with increasing incubation time. Beyond a specific incubation time of the assay (3 min for EA200; 5 min for EA100; 1.8 min for EA70, 30 min EA 40), the fluorescence intensity of the solution remained constant, indicating that all of the PPi had been hydrolyzed by the enzyme. No further fluorescence recovery was observed.
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