A new fluorescent assay for sialyltransferase.
A new fluorescent assay for the sialyltransferase reaction was established. After incubation of the sialyltransferase reaction, the sialyloligosaccharide obtained was treated by acid hydrolysis, and then the NeuAc that was released was labeled with 1,2-diamino-4,5-methylenedioxibenzene. The fluorescent-labeled NeuAc could be estimated by HPLC (excitation: 373 nm; emission: 448 nm) and a Lineweaver-Burk plot could be plotted with the data from this analysis.
Kajihara
Y
Y
Department of System Function, Faculty of Science, Yokohama City University, Japan. kajihara@yokohama-cu.ac.jp
Kamitani
T
T
Sakakibara
T
T
D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes / Q000737:chemistry; D006868:Hydrolysis; D019158:N-Acetylneuraminic Acid / Q000032:analysis; D009844:Oligosaccharides / Q000737:chemistry; D010655:Phenylenediamines / Q000737:chemistry; D012680:Sensitivity and Specificity; D012799:Sialyltransferases / Q000032:analysis*
D005456:Fluorescent Dyes; D009844:Oligosaccharides; D010655:Phenylenediamines; C016735:sialooligosaccharides; C058886:1,2-diamino-4,5-methylenedioxybenzene; D012799:Sialyltransferases; D019158:N-Acetylneuraminic Acid