A sensitive fluorimetric assay for pyruvate.
Anal Biochem
; 396(1): 146-51, 2010 Jan 01.
Article
em En
| MEDLINE
| ID: mdl-19748474
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 11 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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Bioensaio
/
Ácido Pirúvico
/
Fluorometria
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
En
Ano de publicação:
2010
Tipo de documento:
Article