A1 Refereed original research article in a scientific journal
Synthesis of fluorescent oligosaccharides for covalent attachment to living cells
Authors: Gahmberg CG, Tolvanen M
Publication year: 1988
Journal: Analytical Biochemistry
Journal name in source: Analytical biochemistry
Journal acronym: Anal Biochem
Volume: 170
Issue: 2
First page : 520
Last page: 7
ISSN: 0003-2697
DOI: https://doi.org/10.1016/0003-2697(88)90667-7
Abstract
Asparagine-linked oligosaccharides were liberated from glycoproteins by hydrazinolysis. The treatment resulted in de-N-acetylation of the amino sugars. After isolation of the oligosaccharides free amino groups were labeled with fluorescein isothiocyanate and remaining amino groups reacetylated. The fluorescent oligosaccharides were used to label living cells. They were converted to hydrazine derivatives and covalently attached to cell surface oligosaccharides, which had been treated with periodate or neuraminidase and galactose oxidase. This enabled the visualization of the attached oligosaccharides at the external aspect of the plasma membrane by fluorescence microscopy.
Asparagine-linked oligosaccharides were liberated from glycoproteins by hydrazinolysis. The treatment resulted in de-N-acetylation of the amino sugars. After isolation of the oligosaccharides free amino groups were labeled with fluorescein isothiocyanate and remaining amino groups reacetylated. The fluorescent oligosaccharides were used to label living cells. They were converted to hydrazine derivatives and covalently attached to cell surface oligosaccharides, which had been treated with periodate or neuraminidase and galactose oxidase. This enabled the visualization of the attached oligosaccharides at the external aspect of the plasma membrane by fluorescence microscopy.
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