A spectrophotometric collagenase assay

Anal Biochem. 1986 Dec;159(2):390-5. doi: 10.1016/0003-2697(86)90358-1.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autoanalysis
  • Hydrogen-Ion Concentration
  • Microbial Collagenase / analysis*
  • Rosaniline Dyes
  • Spectrophotometry

Substances

  • Rosaniline Dyes
  • Coomassie brilliant blue R
  • Microbial Collagenase