ASSAY OF PROTEOLYTIC-ENZYMES BY THE FLUORESCENCE POLARIZATION TECHNIQUE

被引：69
作者：
MAEDA, H
机构：
[1] Department of Microbiology, Kumamoto University Medical School, Kumamoto
来源：
ANALYTICAL BIOCHEMISTRY | 1979年 / 92卷 / 01期
关键词：
D O I：
10.1016/0003-2697(79)90649-3
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Assay of proteolytic enzymes by the fluorescence polarization method is described in which fluorescein isothiocyanate-conjugated proteins are used as substrates. Hydrolysis of the natural substrates is accompanied by a decrease in the fluorescence polarization values reflecting time and also by an enzyme concentration-dependent decrease in the overall molecular weight of the substrates. Trypsin activity as low as 0.05 μg/ml at 20°C is detected at a fluorochrome concentration of 10 to 20 pmol/ml. In this method, no separation procedure or further reaction for quantitation is necessary. Thus, using a computerized instrument, this method is extremely sensitive, simple, and rapid. © 1979.
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页码：222 / 227
页数：6
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