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An improved method for assay of cholesterol 7 alpha-hydroxylase activity.
- Source :
-
Analytical biochemistry [Anal Biochem] 1986 Oct; Vol. 158 (1), pp. 228-32. - Publication Year :
- 1986
-
Abstract
- An improved assay method for cholesterol 7 alpha-hydroxylase which is accurate, sensitive, and yet still simple is described. The method consists of two parts: the first part is cholesterol 7 alpha-hydroxylation in liver microsomes utilizing cholesterol in situ as the substrate, and the second part is conversion of the product, 7 alpha-hydroxycholesterol, into 7 alpha-hydroxy-4-cholesten-3-one having an intense absorption at 240 nm by the action of cholesterol oxidase. The converted sterol is then analyzed by high-performance liquid chromatography. During the second enzyme reaction, the first enzyme reaction is halted and further metabolism of the product is prevented. In consequence, the method had more than 10-fold increase in the sensitivity than the previous one.
Details
- Language :
- English
- ISSN :
- 0003-2697
- Volume :
- 158
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Analytical biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 3799966
- Full Text :
- https://doi.org/10.1016/0003-2697(86)90613-5