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

  • 佐野 紀代子
    東京医科歯科大学医学部付属病院検査部生化学
  • 長 裕子
    東京医科歯科大学医学部付属病院検査部生化学
  • 保崎 清人
    東京医科歯科大学医学部付属病院検査部生化学

書誌事項

タイトル別名
  • Electrophoretic separation of γ-glutamyl transpeptidase isoenzymes on cellulose acetate “Cellogel”
  • I. Basic investigation on the method
  • I.基礎的検討

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説明

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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