A rapid method for assaying the metabolism of 7-ethoxyresorufin by microsomal subcellular fractions

The formation of resorufin from 7-ethoxyresorufin by microsomal mixed-function oxidase may be assayed rapidly by precipitating the protein in incubation mixtures with methanol and measuring the fluorescence of resorufin in the supernatant after centrifugation. Conditions are described for minimizing...

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Bibliographic Details
Published inAnalytical biochemistry Vol. 107; no. 1; pp. 150 - 155
Main Authors Pohl, Roberta J., Fouts, James R.
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 01.09.1980
Subjects
Animals
Benzoflavones - pharmacology
beta-Naphthoflavone
Cytochrome P-450 CYP1A1
Drug Stability
Edetic Acid
Male
Microsomes - metabolism
Oxazines - analysis
Oxazines - metabolism
Oxidoreductases - metabolism
Rats
Serum Albumin, Bovine
Spectrometry, Fluorescence
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Summary:The formation of resorufin from 7-ethoxyresorufin by microsomal mixed-function oxidase may be assayed rapidly by precipitating the protein in incubation mixtures with methanol and measuring the fluorescence of resorufin in the supernatant after centrifugation. Conditions are described for minimizing the effects of the instability of resorufin and of inhibition by the product. Higher activity and linear plots of microsomal protein concentration versus activity are obtained by including bovine serum albumin or ethylenediamine-tetraacetate in the incubation mixture.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:0003-2697
1096-0309
DOI:10.1016/0003-2697(80)90505-9