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A novel highly sensitive electrophoretic method for the detection and quantification of proteinase activity
Author(s) -
Poulsen Otto M.,
Hau Jann
Publication year - 1986
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150070404
Subject(s) - electrophoresis , agarose , chromatography , substrate (aquarium) , chemistry , penetration (warfare) , hydrolysis , biochemistry , biology , ecology , operations research , engineering
A novel highly sensitive method for the detection and quantification of proteinase activity is presented. The proteinases were forced electrophoretically into the substrate (sodium caseinate) containing agarose gel. The formation of enzyme‐substrate complexes retarded the migration of the proteinases in the electric field and decreased the overall mobility of the enzymes. The proteinases were released as a result of the degradation of the substrate, and consequently the height of the obtained rocket shaped zones of hydrolysis were proportional to the concentration of proteinases in the samples. The electrophoretic method accelerated penetration of the substrate‐containing gel by the proteinases and subsequently resulted in a 100‐fold increase in sensitivity as compared to the radial diffusion method, depending entirely on diffusion.