A modified colorimetric method of gelatinolytic assay using bacterial collagenase type II as a model
A modified colorimetric method of gelatinolytic assay using bacterial collagenase type II as a model
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DOI：
10.1016/j.ab.2012.09.036
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发表时间：
2013-02-15
影响因子：
2.9
通讯作者：
Chomdej, Siriwadee
中科院分区：
文献类型：
--
作者：
Osathanunkul, Maslin;Buddhachat, Kittisak;Chomdej, Siriwadee
用微波加热的考马斯亮蓝（CBB）作为染色染料测定明胶分解活性。使用加热的CBB测定与细菌胶原酶孵育后剩余的明胶量，得到可见的蓝色颗粒。加入二甲亚砜以溶解染料，并测量600 nm处的吸光度以检测明胶水平（最高10 μ g），胶原酶量的检测限为50 ng。这种方法是快速，简单，和经济的目的，筛选药物具有抑制活性的胶原酶。(C)2012 Elsevier Inc. All rights reserved.
Coomassie brilliant blue (CBB) heated by microwave was used as a staining dye for measuring gelatinolytic activity. The quantity of gelatin remaining after incubation with bacterial collagenase was determined using the heated CBB, resulting in visible blue pellets. Dimethyl sulfoxide was added to dissolve the dye and measurement of the absorbance at 600 nm was done to detect the level of gelatin (up to 10 mu g), with the limit of detection for the amount of collagenase at 50 ng. This approach is rapid, simple, and economic for the purpose of screening for pharmaceutical agents that possess inhibitory activity on collagenase. (C) 2012 Elsevier Inc. All rights reserved.