Acyl-coenzyme A:cholesterol acyltransferase assay: Silica gel column separation of reaction products
Identifieur interne : 000182 ( France/Analysis ); précédent : 000181; suivant : 000183Acyl-coenzyme A:cholesterol acyltransferase assay: Silica gel column separation of reaction products
Auteurs : Magali Chautan [France] ; Elise Termine [France] ; Gilles Nalbone [France] ; Huguette Lafont [France]Source :
- Analytical Biochemistry [ 0003-2697 ] ; 1988.
English descriptors
Abstract
Acyl-coenzyme A:cholesterol acyltransferase (ACAT) assays are usually performed by incubation of the enzyme with a labeled substrate followed by thin-layer chromatography separation and subsequent quantification of cholesteryl esters (CE) formed. Herein, a method is described for rapid separation of CE from other lipids, by elution from a silica gel column with a solvent mixture of petroleum ether/diethyl ether (98:2, v/v). Silica gel column chromatography is reliable and more rapid and safer than TLC. The best results were obtained when the reaction was stopped by Dole extraction followed by CE separation on a silica gel column. Assays for ACAT from rat intestinal microsomes showed that the specific activity values obtained using this method were reproducible and in good agreement with those obtained by conventional TLC method.
Url:
DOI: 10.1016/0003-2697(88)90210-2
Affiliations:
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Herein, a method is described for rapid separation of CE from other lipids, by elution from a silica gel column with a solvent mixture of petroleum ether/diethyl ether (98:2, v/v). Silica gel column chromatography is reliable and more rapid and safer than TLC. The best results were obtained when the reaction was stopped by Dole extraction followed by CE separation on a silica gel column. 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