FLUORESCEIN ISOTHIOCYANATE-LABELED CASEIN ASSAY FOR PROTEOLYTIC-ENZYMES
FLUORESCEIN ISOTHIOCYANATE-LABELED CASEIN ASSAY FOR PROTEOLYTIC-ENZYMES
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DOI：
10.1016/0003-2697(84)90553-0
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发表时间：
1984-01-01
影响因子：
2.9
通讯作者：
TWINING, SS
中科院分区：
文献类型：
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作者：
TWINING, SS
利用可溶性异硫氰酸荧光素(FITC)标记酪蛋白，建立了一种简单、廉价、灵敏的蛋白水解酶分析方法。酪蛋白与FITC反应生成荧光素硫代氨基甲酰衍生物。该底物被胰酶、胰凝乳蛋白酶、弹性蛋白酶、枯草杆菌酶和热裂解酶以线性的时间依赖方式切割。这些酶可以用这种分析方法在ng和亚ng范围内进行测量。该检测方法重复性好，空白含量低，并使用酪蛋白，它类似于大多数蛋白酶的天然底物。
A simple inexpensive sensitive protease assay was developed using soluble fluorescein isothiocyanate (FITC)-labeled casein. Casein was reacted with FITC to form the fluorescein thiocarbamoyl derivative. This substrate is cleaved by trypsin, chymotrypsin, elastase, subtilisin and thermolysin in a linear time-dependent manner. These enzymes can be measured in the ng and sub-ng range using this assay. The assay is reproducible, has a low blank and uses casein, which resembles natural substrates of most proteases.