A spectrophotometric collagenase assay.
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.
D001322:Autoanalysis; D006863:Hydrogen-Ion Concentration; D003012:Microbial Collagenase / Q000032:analysis*; D012394:Rosaniline Dyes; D013053:Spectrophotometry
D012394:Rosaniline Dyes; C024757:Coomassie brilliant blue R; D003012:Microbial Collagenase