A simple colorimetric assay method for pyrophosphate in the presence of a 1000-fold excess of orthophosphate: application of the method to the study of pyrophosphate metabolism in mitochondria.
A sensitive colorimetric method for the assay of inorganic pyrophosphate with excess of orthophosphate is described. The principle of this method lies in the formation of phosphomolybdate and PPi-molybdate complexes with subsequent extraction of the phosphomolybdate complex by organic solvents and reduction of the PPi-molybdate complex by dithiothreitol and Eikonogen. The sensitivity of the method was from 5 to 120 nmol of PPi in a 2.0-ml sample.
Mansurova
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A. N. Belozersky Laboratory of Molecular Biology, Moscow State University, USSR.
Deryabina
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D000818:Animals; D002138:Calibration; D003124:Colorimetry; D011756:Diphosphates / Q000032:analysis / Q000378:metabolism*; D004229:Dithiothreitol; D005459:Fluorides; D008928:Mitochondria / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000378:metabolism; D008982:Molybdenum / Q000378:metabolism; D010084:Oxidation-Reduction; D010710:Phosphates / Q000032:analysis*; D051381:Rats
D011756:Diphosphates; D010710:Phosphates; C044659:molybdate; D008982:Molybdenum; D005459:Fluorides; D004229:Dithiothreitol