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A rapid sensitive collagenase assay

โœ Scribed by M.T. Gisslow; B.C. McBride

Publisher
Elsevier Science
Year
1975
Tongue
English
Weight
448 KB
Volume
68
Category
Article
ISSN
0003-2697

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โœฆ Synopsis

A rapid, sensitive collagenase assay has been developed using 14C-acetylated collagen as a substrate. Acid-soluble calfskin collagen was labeled with [1-14C]acetic anhydride at pH 8. The acetylated collagen had a specific activity of 6.25 ร— l0 ~ dpm/mg protein. Collagen was not denatured as evidenced by its resistance to nonspecific proteolysis and sensitivity to bacterial collagenase. Polyacrylamide gel electrophoresis of the acetylated protein showed that the radioactivity was present in the three bands corresponding to the a,/3, and 3' components of collagen. The rate of release of 14C from labeled collagen by Clostridium histolyticum collagenase was proportional to enzyme and substrate concentration.

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