High-efficiency labeling of sialylated glycoproteins on living cells
Nature methods, 2009•nature.com
We describe a simple method for efficiently labeling cell-surface sialic acid–containing
glycans on living animal cells. The method uses mild periodate oxidation to generate an
aldehyde on sialic acids, followed by aniline-catalyzed oxime ligation with a suitable tag.
Aniline catalysis dramatically accelerates oxime ligation, allowing use of low concentrations
of aminooxy-biotin at neutral pH to label the majority of cell-surface sialylated glycoproteins
while maintaining high cell viability.
glycans on living animal cells. The method uses mild periodate oxidation to generate an
aldehyde on sialic acids, followed by aniline-catalyzed oxime ligation with a suitable tag.
Aniline catalysis dramatically accelerates oxime ligation, allowing use of low concentrations
of aminooxy-biotin at neutral pH to label the majority of cell-surface sialylated glycoproteins
while maintaining high cell viability.
Abstract
We describe a simple method for efficiently labeling cell-surface sialic acid–containing glycans on living animal cells. The method uses mild periodate oxidation to generate an aldehyde on sialic acids, followed by aniline-catalyzed oxime ligation with a suitable tag. Aniline catalysis dramatically accelerates oxime ligation, allowing use of low concentrations of aminooxy-biotin at neutral pH to label the majority of cell-surface sialylated glycoproteins while maintaining high cell viability.
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