Abstract
An improved assay method for cholesterol 7α-hydroxylase which is accurate, sensitive and yet still simple is described. The method consists of two parts: the first part is cholesterol 7α-hydroxylation in liver microsomes utilizing cholesterol in situ as the substrate, and the second part is conversion of the product, 7α-hydroxycholesterol, into 7α-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.
| Original language | English |
|---|---|
| Pages (from-to) | 228-232 |
| Number of pages | 5 |
| Journal | Analytical Biochemistry |
| Volume | 158 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Oct 1986 |
| Externally published | Yes |
All Science Journal Classification (ASJC) codes
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology