Literature DB >> 28359787

Tips on the analysis of phosphatidic acid by the fluorometric coupled enzyme assay.

Azam Hassaninasab1, Gil-Soo Han1, George M Carman2.   

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.
Copyright © 2017 Elsevier Inc. All rights reserved.

Entities:  

Keywords:  Coupled enzyme assay; Fluorometric assay; Phosphatidic acid; Phosphatidic acid phosphatase; Yeast

Mesh：

Substances：

Year:  2017        PMID： 28359787      PMCID： PMC5425298          DOI： 10.1016/j.ab.2017.03.020

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


  19 in total

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