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This method is used in assaying inorganic phosphate in the presence of ATP. Exactly 5.0 ml of an assay mixture containing water, ammonium molybdate reagent (4% ammonium molybdate in molybdate preservative), and reductant catalyst in a 1: 2: 3 ratio, espectively, is added to 1.0 ml of the sample. After a waiting time of 2 min, 0.5 ml of color developer is added while the solution is mixed on a Vortex mixer.The absorbance is then read at any time between 10 and 40 min on a spectrophotometer. Absorbance can be read at any wavelength between 550 and 720 nm, with maximum sensitivity at 720 nm (the ratio of theabsorbance readings at 550, 600, and 720 nm is 0.5 :0.7: 1, respectively).The entire assay is done at room temperature (25”C), though it ay also be run at 0°C.

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