J. Lipid Res. Acyl Labeled PIP's available August 1, 2008
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Journal of Lipid Research, Vol 22, 720-724, Copyright © 1981 by Lipid Research, Inc.

ARTICLES

Measurement of cholesterol 7 alpha-hydroxylase activity with selected ion monitoring

A Sanghvi, E Grassi, C Bartman, R Lester, M Galli Kienle and G Galli

Formation of 7 alpha-hydroxycholesterol by rat liver microsomes was quantitated using a gas chromatograph-mass spectrometer operated in selected ion monitoring mode. This procedure measures the mass of 7 alpha-hydroxycholesterol formed and does not require addition of labeled cholesterol to monitor the reaction, since endogenous microsomal cholesterol serves as substrate for the enzyme cholesterol 7 alpha-hydroxylase. The method requires only the extraction of lipids from microsomal incubations and injection of a small fraction of this extract into the apparatus. Microsomes from normal rat livers incubated for different periods were found to yield increased 7 alpha- hydroxycholesterol with time.
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Copyright © 1981 by the American Society for Biochemistry and Molecular Biology.