A modified colorimetric method of gelatinolytic assay using bacterial collagenase type II as a model

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 di...

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محفوظ في:
التفاصيل البيبلوغرافية
المؤلفون الرئيسيون: Maslin Osathanunkul, Kittisak Buddhachat, Siriwadee Chomdej
التنسيق: دورية
منشور في: 2018
الوصول للمادة أونلاين:https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84870695160&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/48106
الوسوم: إضافة وسم
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المؤسسة: Chiang Mai University
الوصف
الملخص: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 μg), 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. © 2012 Elsevier Inc. All rights reserved.