Activity staining of pectinesterase on polyacrylamide gels after acidic or sodium dodecyl sulfate electrophoresis

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Abstract
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 β‐naphthyl acetate (β‐NA). The release of β‐naphthol (at 322 nm) from β‐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 β‐NA. This fast and sensitive method can be used for enzyme purification and characterization.