Nethery A, Lyons J G, O'Grady R L
Anal Biochem. 1986 Dec;159(2):390-5. doi: 10.1016/0003-2697(86)90358-1.
A quantitative collagenase assay using Coomassie blue staining and microtiter spectrophotometry is described. Collagen is gelled and dried onto the bottom of microwells as substrate, washed, incubated with samples, washed again, and then stained. Absorbance at 590 nm increases linearly with increasing amounts of collagen in the range 5-40 micrograms. Bacterial and mammalian collagenases can be detected within 2 h, and 10 ng of bacterial collagenase may be detected in 16 h. For simple screening applications, activity may be detected by eye. The assay is safe, simple, fast, economical, and sensitive.
本文描述了一种使用考马斯亮蓝染色和微量滴定分光光度法的定量胶原酶测定方法。将胶原蛋白凝胶化并干燥在微孔底部作为底物,洗涤后与样品孵育,再次洗涤,然后染色。在5-40微克范围内,590纳米处的吸光度随胶原蛋白量的增加呈线性增加。细菌和哺乳动物胶原酶可在2小时内检测到,16小时内可检测到10纳克细菌胶原酶。对于简单的筛选应用,可通过肉眼检测活性。该测定方法安全、简单、快速、经济且灵敏。