Fluorescent staining of sialidases in polyacrylamide gel electrophoresis and ultrathin-layer isoelectric focusing.
Polyacrylamide gels were stained with the sialidase substrate 2'-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid showing the activity of Vibrio cholerae and Clostridium sordellii sialidases in the gels after electrophoresis. With this fluorogenic method minimum sialidase activities of 5 microU could be determined. The sensitivity of this staining is about 10,000-fold higher compared to protein-staining with Coomassie brilliant blue. For the visualization of other proteins than sialidases the specific sialidase staining could be followed by a protein-staining method in the same gel.
Berg
W
W
Gutschker-Gdaniec
G
G
Schauer
R
R
D055598:Chemical Phenomena; D002621:Chemistry; D003013:Clostridium / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes; D006923:Hymecromone / Q000031:analogs & derivatives; D007525:Isoelectric Focusing / Q000379:methods; D009439:Neuraminidase / Q000302:isolation & purification*; D013050:Spectrometry, Fluorescence; D013194:Staining and Labeling; D014468:Umbelliferones; D014734:Vibrio cholerae / Q000201:enzymology
D005456:Fluorescent Dyes; D014468:Umbelliferones; D006923:Hymecromone; C021224:2'-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid; D009439:Neuraminidase