Activity staining of pectinesterase on polyacrylamide gels after acidic or sodium dodecyl sulfate electrophoresis.
Pectinesterase (PE), from commercial orange peels or ammonium sulfate fractionation (50-80% saturation) of pea pods, was detected on polyacrylamide gels after native acidic polyacrylamide gel electrophoresis (PAGE) or sodium dodecyl sulfate (SDS)-PAGE by using the synthetic substrate beta-naphthyl acetate (beta-NA). The release of beta-naphthol (at 322 nm) from beta-NA was proportional to PE activity. The PE activity bands on polyacrylamide gels after native acidic PAGE or SDS-PAGE were stained with a combination of tetrazotized o-dianisidine and beta-NA. This fast and sensitive method can be used for enzyme purification and characterization.
Hou
W C
WC
Institute of Botany, Academica Sinica, Nankang, Taipei, Taiwan, ROC.
Lin
Y H
YH
D000180:Acrylic Resins; D002265:Carboxylic Ester Hydrolases / Q000032:analysis*; D004396:Coloring Agents; D003962:Dianisidine; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D009596:Nitrophenols / Q000378:metabolism; D018532:Pisum sativum / Q000201:enzymology*; D010368:Pectins; D012430:Ruthenium Red; D012967:Sodium Dodecyl Sulfate; D013194:Staining and Labeling / Q000379:methods; D013379:Substrate Specificity
D000180:Acrylic Resins; D004396:Coloring Agents; D009596:Nitrophenols; C016680:polyacrylamide gels; D012430:Ruthenium Red; D012967:Sodium Dodecyl Sulfate; C008642:4-nitrophenyl acetate; D010368:Pectins; D002265:Carboxylic Ester Hydrolases; C022323:pectinesterase; D003962:Dianisidine