A versatile microassay for elastase using succinylated elastin

被引:29
作者
Rao, SK [1 ]
Mathrubutham, M [1 ]
Karteron, A [1 ]
Sorensen, K [1 ]
Cohen, JR [1 ]
机构
[1] PIERCE CHEM CO,ROCKFORD,IL
关键词
D O I
10.1006/abio.1997.2223
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We have developed a rapid, versatile, and sensitive elastase assay that is based on the measurement of primary amines that ape exposed due to enzymatic degradation of proteins, using succinylated elastin as the substrate for elastase. After incubation with elastase the degree of digestion is determined with trinitrobenzene sulfonic acid. The assay is rapid and sensitive, detecting elastase down to 1 ng/ml, and is linear up to enzyme concentrations of 10 mu g/ml. The assay is carried out in microtiter plate wells and therefore offers the potential for assaying numerous samples of small volume. The use of succinylated elastin shows specificity for elastase over the control protease, trypsin. This assay is also versatile because it can be applied to samples such as cell culture supernatants, blood plasma, tissue biopsies, and tissue homogenates. (C) 1997 Academic Press.
引用
收藏
页码:222 / 227
页数:6
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