Method for the detection of substrate specific protease activity by using cellulose acetate membrane as a absorbant of substrate.
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概要
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We present here a rapid method for determining substrate specificities of protease activities by using two-dimensional polyacrylamide gel electrophoresis and cellulose acetate membranes. Proteolytic active exzymes were satisfactorily separated by the two-dimensional polyacrylamide gel electrophoresis. Determining of the substrate specificities of protease activities were accomplished by using cellulose acetate membranes as absorbent of substrates. Two sheets of cellulose acetate membranes containing different substrates respectively were placed on the both sides of the thin gel layer. During incubation, enzyme activities were transferred to the membranes from the gel layer. After incubation, the membranes were stripped from the gel layer and each substrate hydrolyzing activity was detected. Comparing the zymograms, substrate specificities of proteolytic active spots were easily determined. This method is sensitive and rapid, allowing us to determine the precise substrate specificities of protease activities from only one gel layer.
- 日本電気泳動学会の論文
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