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Measurement of phosphate concentration in the presence of very labile phosphate esters
Authors:Russell J. M. Lane  Nicholas J. Watmough
Affiliation:Department of Neurochemistry, Regional Neurological Centre, Newcastle General Hospital, Westgate Road, Newcastle upon Tyne, NE4 6BE, England
Abstract:There are a number of methods available for the measurement of phosphate ion concentration, which may be used when moderately labile phosphate esters such as ATP are present in low concentration. However, the highly acidic conditions usually employed make these unsuitable when very labile esters such as phosphocreatine are present. A method in which the phosphomolybdate complex is developed under mildly acidic conditions, using high molybdate concentrations to counteract the reduced assay sensitivity at high pH, is described. The assay is linear in the range 5-300 microM phosphate, and micromolar concentrations of phosphate can be reliably measured in the presence of millimolar phosphocreatine.
Keywords:phosphate assay   ATP   phosphocreatine   ATPases
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