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Improved detection of amylase activity by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis with copolymerized starch
Author(s) -
Martínez Tomás F.,
Alarcón Francisco J.,
DíazLópez Manuel,
Moyano Francisco J.
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(20000801)21:14<2940::aid-elps2940>3.0.co;2-s
Subject(s) - sodium dodecyl sulfate , amylase , chromatography , chemistry , polyacrylamide gel electrophoresis , molecular mass , hydrolysis , starch , gel electrophoresis , electrophoresis , sodium , polyacrylamide , capillary electrophoresis , biochemistry , enzyme , polymer chemistry , organic chemistry
An improved method, based on sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) for detection of amylase activity is described. This method will allow better characterization of certain amylases than that obtained by the Davis technique. The main features of the technique are: (i) identification of amylase bands and molecular mass determination are possible in the same gel; (ii) the hydrolysis of copolymerized substrate during electrophoretic separation is prevented using very low temperatures instead of inactivating agents such as chelating agents; and (iii) the technique is applicable to reveal amylase activity in a wide range of biological samples. The method is not useful for enzymes sensitive to SDS and for high molecular mass amylases.