A versatile microassay for elastase using succinylated elastin
A versatile microassay for elastase using succinylated elastin
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DOI：
10.1006/abio.1997.2223
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发表时间：
1997-08-01
影响因子：
2.9
通讯作者：
Cohen, JR
中科院分区：
文献类型：
--
作者：
Rao, SK;Mathrubutham, M;Cohen, JR
我们已经开发了一种快速，通用，和敏感的弹性蛋白酶测定，这是基于测量的伯胺，猿暴露由于蛋白质的酶促降解，使用琥珀酰化弹性蛋白作为弹性蛋白酶的底物。与弹性蛋白酶孵育后，用三硝基苯磺酸测定消化程度。该测定法是快速和灵敏的，检测低至1 ng/ml的弹性蛋白酶，并且在高达10 μ g/ml的酶浓度下呈线性。该测定在微量滴定板威尔斯孔中进行，因此提供了测定大量小体积样品的可能性。琥珀酰化弹性蛋白的使用显示出对弹性蛋白酶的特异性超过对照蛋白酶胰蛋白酶。该测定也是通用的，因为它可以应用于样品，如细胞培养上清液、血浆、组织活检和组织匀浆。(C)北京：科学出版社.
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.