Use of fluorescamine-labeled casein as a substrate for assay of proteinases

Kazuhiro Sogawa, Kenji Takahashi

    研究成果: Article査読

    32 被引用数 (Scopus)

    抄録

    Conditions have been investigated for the use of fluorescamine-labeled casein as a substrate for fluorometric assay of proteinases. Fluorescamine-labeled casein can be prepared simply by mixing solutions of casein and fluorescamine at pH 8.0 and used without removal of the excess reagent or its hydrolysis product. The fluorescence of the labeled casein and its enzymatic digest is moderately stable in the range of pH 7.0 to 10.0. Activities can be determined by measuring the fluorescence of the hydrolysis products soluble in 0.1 M trichloroacetie acid solution at pH 4.0 after adjusting the pH of the acid-soluble fraction to 7.7. This method is suited for assay of proteinases active at neutral to slightly alkaline pH values, and is capable of quantitating about 0.05 μg of trypsin or 0.5 μg of α-chymotrypsin or papain. The assay can be done in the presence of large amounts of contaminating amino acid, protein and/or exopeptidases which may interfere with the ordinary assay of proteinases.

    本文言語English
    ページ(範囲)1783-1787
    ページ数5
    ジャーナルJournal of biochemistry
    83
    6
    DOI
    出版ステータスPublished - 1978 6

    ASJC Scopus subject areas

    • 生化学
    • 分子生物学

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