Improved detection of amylase activity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with copolymerized starch

Electrophoresis. 2000 Aug;21(14):2940-3. doi: 10.1002/1522-2683(20000801)21:14<2940::AID-ELPS2940>3.0.CO;2-S.

Abstract

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.

MeSH terms

  • Amylases / analysis*
  • Animals
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Starch

Substances

  • Starch
  • Amylases