Visualization of acid phosphatase activity on nitrocellulose filters following electroblotting of polyacrylamide gels.
A method for visualizing acid phosphatase isoenzymes by activity staining on nitrocellulose filters after electroblotting of proteins fractionated on nondenaturing polyacrylamide gels is described. Reproducible results were obtained when 25 mM Tris-192 mM glycine was used as the transfer buffer instead of 0.7% acetic acid, 50 mM sodium acetate, pH 4, or 0.14 M acetic acid--0.35 M beta-alanine, pH 4.3. Dot-blot analysis of banana fruit extracts on nitrocellulose filters revealed that a minimum of 5 x 10(-3) units (nmol p-nitrophenyl phosphate hydrolyzed g-1.h-1) of acid phosphatase activity can be detected. This method can be suitable for screening a large number of biological samples for monitoring acid phosphatase activity.
Kanellis
A K
AK
Department of Horticulture, University of Maryland, College Park 20742.
Solomos
T
T
Mattoo
A K
AK
D000135:Acid Phosphatase / Q000032:analysis*; D003101:Collodion; D004591:Electrophoresis, Polyacrylamide Gel; D005374:Filtration; D007527:Isoenzymes / Q000032:analysis; D010944:Plants / Q000201:enzymology; D013194:Staining and Labeling
D007527:Isoenzymes; D003101:Collodion; D000135:Acid Phosphatase