Elsevier

Analytical Biochemistry

Volume 102, Issue 1, February 1980, Pages 196-202
Analytical Biochemistry

Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates

https://doi.org/10.1016/0003-2697(80)90338-3Get rights and content

Abstract

A new technique is described for the electrophoretic analysis of plasminogen activators in sodium dodecyl sulfate-polyacrylamide gels containing copolymerized plasminogen and gelatin. The method depends upon the fact that the zymogen and gelatin, when incorporated into the polyacrylamide matrix at the time of casting, are retained during subsequent electrophoresis of enzyme samples, and serve as satisfactory sequential, in situ substrates for the localization of plasminogen activator bands by negative staining. The nonionic detergent, Triton X-100, is used to remove sodium dodecyl sulfate and restore enzyme activity. The method can be used to detect as little as 1 mU of urokinase and effectively distinguishes between melanoma- and urokinase-type plasminogen activators. Plasminogen-independent proteases are detected by omission of plasminogen from the gel.

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