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Assay of long-chain acyl-CoAs in a complex reaction mixture.

作者信息

Juguelin H, Cassagne C

出版信息

Anal Biochem. 1984 Nov 1;142(2):329-35. doi: 10.1016/0003-2697(84)90472-x.

DOI:10.1016/0003-2697(84)90472-x
PMID:6528971
Abstract

A method has been developed which allows the quantitative analysis of labeled or unlabeled acyl-CoAs in complex reaction mixtures. The method is based on (a) a quantitative solubilization of acyl-CoAs and lipids, directly in the reaction vessel, by 0.05 M Tris-HCl, pH 7.5/CHCl3/CH3OH (1/3/3, v/v/v); (b) monodimensional TLC of aliquots of the whole reaction mixture, resolving malonyl-CoA, acetyl-CoA, long-chain acyl-CoAs, polar lipids and neutral lipids plus free fatty acids; and (c) quantitation by TLC densitometry and/or TLC radiochromatography. All fractions--and particularly long chain-acyl-CoAs--can then be analyzed for distribution and label of fatty acyl moieties.

摘要

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