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Literature DB >> 19748474

A sensitive fluorimetric assay for pyruvate.

Aiping Zhu¹, Roberto Romero, Howard R Petty.

Abstract

A sensitive and specific fluorimetric assay for the determination of pyruvate is reported here. This assay is based on the oxidation of pyruvate in the presence of pyruvate oxidase. Hydrogen peroxide generated by pyruvate oxidase reacts with nonfluorescent Amplex Red at a 1:1 stoichiometry to form the fluorescent product, resorufin. The assay is optimized with respect to pH of reaction buffer, enzyme concentration, dye concentration, and the time course. The usefulness of the assay is demonstrated by the accurate measurement of intracellular and extracellular pyruvate concentrations. The limit of detection of the assay is 5 nM.

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Year: 2009 PMID： 19748474 PMCID： PMC2790020 DOI： 10.1016/j.ab.2009.09.017

Source DB: PubMed Journal: Anal Biochem ISSN： 0003-2697 Impact factor: 3.365

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