J. Lipid Res. Acyl Labeled PIP's available August 1, 2008
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Journal of Lipid Research, Vol 16, 92-96, Copyright © 1975 by Lipid Research, Inc.

ARTICLES

Isotope derivative assay of microsomal cholesterol 7 alpha-hydroxylase

S Shefer, G Nicolau and EH Mosbach

A rapid method was developed to measure cholesterol 7 alpha-hydroxylase activity of hepatic microsomes by the direct determination of the mass of 7 alpha-hydroxycholesterol formed. The method is based on the quantitative acetylation of the incubation mixture with [-3H]acetic anhydride and the separation of the biosynthetic 7 alpha- hydroxycholesterol as its diacetate by thin-layer chromatography on alumina. Amounts of 7 alpha-hydroxycholesterol as low as 0.1 nmole could be measured. A comparison of the proposed isotope derivative method with the previously used isotope incorporation method showed that the latter underestimated the enzyme activity by about 20 percent.
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