Application of Microcolumns of Activated Carbon to the Monitoring of Glutathione S-Transferase Activity toward Aliphatic Substrates by Thin-Layer Chromatodensitometry

概要

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The present study was carried out in order to develop a method for monitoring glutathione S-transferase activity toward aliphatic compounds^<2)> in fractions of biological protein samples obtained by column chromagraphy. Microcolumns of activated carbon (8 mm long, 7 mm in diameter, smaller than 200 mesh, 80 mg) were used for the separation of glutathione conjugates, which were formed by incubating an aliphatic substrate with rat liver enzyme solutions fractionated by DEAE- or CM-cellulose column chromatography. Subsequently about 90 sample solutions obtained from the microcolumns were all subjected to thin-layer chromatography. After staining of the chromatograms with Ninhydrin reagent, the line of spots corresponding to the glutathione conjugate was scanned on a densitometer. The enzyme activity curve obtained by densitometry was confirmed to represent the glutathione S-transferase activity by comparing the results with those of conventional spectrophotometric assays employing 1-chloro-2,4-dinitrobenzene as the substrate.
公益社団法人日本薬学会の論文
1982-10-30