An Improved Method for the Determination of Cytoplasmic 3α- and 17β- and Microsomal 3α-Hydroxysteroid Dehydrogenase Activities in Rat Liver
| Autor: | Herbert Schriefers, Edmund Rodney Lax, Ernst Kreuzfelder |
|---|---|
| Rok vydání: | 1979 |
| Předmět: |
Male
3-Hydroxysteroid Dehydrogenases 17-Hydroxysteroid Dehydrogenases Alpha (ethology) Tritium Biochemistry Cofactor Cytosol Sex Factors Species Specificity Animals Centrifugation Carbon Radioisotopes Hydroxysteroid dehydrogenase chemistry.chemical_classification biology Chemistry Substrate (chemistry) Rats Kinetics Enzyme Liver Cytoplasm Isotope Labeling Microsomes Liver Microsome biology.protein Female |
| Zdroj: | Hoppe-Seyler´s Zeitschrift für physiologische Chemie. 360:897-904 |
| ISSN: | 0018-4888 |
| DOI: | 10.1515/bchm2.1979.360.2.897 |
| Popis: | This paper described a modified method for the radiometric determination of hydroxysteroid dehydrogenase activities in rat liver. The principle advantages of this method are the improved precision and a radical reduction in the time involved in performing the assay. The procedure comprises the following steps: incubation of 14C-labelled substrate with coenzyme and cell fraction under optimized conditions; termination of the reaction by addition of organic solvent containing a defined amount of 3H-labelled reaction product; removal of precipitated protein and coenzyme by centrifugation; paper chromatographic isolation of the product; direct quantitation of 14C activity in the product zone of the paper chromatogram. The assay systems have been applied to elucidate and quantitate sex and strain differences in the activities of the above enzymes in Chbb/THOM and Sprague-Dawley rats. |
| Databáze: | OpenAIRE |
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