A two-step purification of cytochrome P-450 from adult pig testis by pregnenolone affinity column chromatography
✍ Scribed by Masahiro Kuwada
- Publisher
- John Wiley and Sons
- Year
- 1999
- Tongue
- English
- Weight
- 141 KB
- Volume
- 13
- Category
- Article
- ISSN
- 0269-3879
No coin nor oath required. For personal study only.
✦ Synopsis
Adult testicular cytochrome P-450 was purified by a two-step procedure utilizing hydroxylapatite and pregnenolone affinity column chromatography. The cytochrome P-450 was determined to have an isoelectric point of 6.43 on analytical isoelectric focusing. The purified cytochrome P-450 was found to be homogeneous and its molecular weight was estimated to be 52,000 on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The carbon monoxide difference spectrum with a peak at 448 nm exhibited the absorption spectrum of a typical cytochrome P-450. A purification of 755x was achieved with a yield of 3.09%.