Improved detection of amylase activity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with copolymerized starch.
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.
Martínez
T F
TF
Departamento de Biología Aplicada, Universidad de Almería, Spain.
Alarcón
F J
FJ
Díaz-López
M
M
Moyano
F J
FJ
D000681:Amylases / Q000032:analysis*; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D013213:Starch
D013213:Starch; D000681:Amylases