Tips on the analysis of phosphatidic acid by the fluorometric coupled enzyme assay.
Anal Biochem
; 526: 69-70, 2017 06 01.
Article
em En
| MEDLINE
| ID: mdl-28359787
ABSTRACT
The fluorometric coupled enzyme assay to measure phosphatidic acid (PA) involves the solubilization of extracted lipids in Triton X-100, deacylation, and the oxidation of PA-derived glycerol-3-phosphate to produce hydrogen peroxide for conversion of Amplex Red to resorufin. The enzyme assay is sensitive, but plagued by high background fluorescence from the peroxide-containing detergent and incomplete heat inactivation of lipoprotein lipase. These problems affecting the assay reproducibility were obviated by the use of highly pure Triton X-100 and by sufficient heat inactivation of the lipase enzyme. The enzyme assay could accurately measure the PA content from the subcellular fractions of yeast cells.
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MEDLINE
Assunto principal:
Ácidos Fosfatídicos
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Saccharomyces cerevisiae
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Octoxinol
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Ensaios Enzimáticos
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Fluorometria
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Lipase Lipoproteica
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article