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 - Jan 1 1986 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology
Cite this
An improved method for assay of cholesterol 7α-hydroxylase activity. / Ogishima, Tadashi; Okuda, Kyuichiro.
In: Analytical Biochemistry, Vol. 158, No. 1, 01.01.1986, p. 228-232.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - An improved method for assay of cholesterol 7α-hydroxylase activity
AU - Ogishima, Tadashi
AU - Okuda, Kyuichiro
PY - 1986/1/1
Y1 - 1986/1/1
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=0023009316&partnerID=8YFLogxK
U2 - 10.1016/0003-2697(86)90613-5
DO - 10.1016/0003-2697(86)90613-5
M3 - Article
C2 - 3799966
AN - SCOPUS:0023009316
VL - 158
SP - 228
EP - 232
JO - Analytical Biochemistry
JF - Analytical Biochemistry
SN - 0003-2697
IS - 1
ER -