Electrophoretic separation of γ-glutamyl transpeptidase isoenzymes on cellulose acetate “Cellogel”

DOI
  • Sano Kiyoko
    東京医科歯科大学医学部付属病院検査部生化学
  • Cho Hiroko
    東京医科歯科大学医学部付属病院検査部生化学
  • Hosaki Seijin
    東京医科歯科大学医学部付属病院検査部生化学

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Other Title
  • セルロースアセテート膜“Ｃｅｌｌｏｇｅｌ”によるγ‐ＧＴＰアイソザイム分画　Ｉ　　基礎的検討
  • I. Basic investigation on the method
  • I.基礎的検討

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A method was described for separation of γ-glutamyl transpeptidase (γ-GTP) isoenzymes by cellulose acetate membrane (Cellogel). Enzyme activity staining was carried out by use of γ-L-glutamyl-α-naphthylamide as substrate and fast blue B as color developing agent. Zymogram on Cellogel membrane was treated with 5% trichloroacetic acid solution and applied for densitometry.<br>The conditions for this method (concentration of substrate, diazo reagent, glycylglycine, pH of buffer, incubation time, etc.) were examined and optimal conditions were proposed.<br>This method was simple and sensitive enough to detect γ-GTP isoenzyme patterns from the normal serum containing 30mU/ml activity. Linear relationship between color development of zymogram and enzyme activity was also obtained from 30mU/ml to 230mU/ml in serum sample and this method, therefore, was available for the routine work as semi-quantitative method.

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