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Activity staining of pectinesterase on polyacrylamide gels after acidic or sodium dodecyl sulfate electrophoresis
Author(s) -
Hou WenChi,
Lin YawHuei
Publication year - 1998
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/elps.1150190515
Subject(s) - polyacrylamide , polyacrylamide gel electrophoresis , chemistry , chromatography , sodium dodecyl sulfate , pectinesterase , sodium , gel electrophoresis , staining , ammonium sulfate , biochemistry , enzyme , biology , pectinase , organic chemistry , polymer chemistry , genetics
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.