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.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Aldehydes / chemistry
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Aniline Compounds / chemistry
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Animals
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Biotin / analogs & derivatives
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Biotin / chemistry
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Biotinylation / methods*
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CHO Cells
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Catalysis
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Cell Line
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Cell Survival
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Cricetinae
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Cricetulus
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Endocytosis
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Flow Cytometry
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Humans
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Leukocyte Common Antigens / chemistry
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Membrane Glycoproteins / chemistry*
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Microscopy, Fluorescence
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N-Acetylneuraminic Acid / analogs & derivatives
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N-Acetylneuraminic Acid / chemistry*
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Oxidation-Reduction
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Oximes / chemistry
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Periodic Acid / chemistry
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Sialyltransferases / chemistry
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Staining and Labeling / methods*
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Streptavidin / chemistry
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beta-D-Galactoside alpha 2-6-Sialyltransferase
Substances
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Aldehydes
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Aniline Compounds
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Membrane Glycoproteins
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Oximes
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Periodic Acid
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Biotin
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Streptavidin
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metaperiodate
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Sialyltransferases
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Leukocyte Common Antigens
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PTPRC protein, human
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N-Acetylneuraminic Acid
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aniline
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beta-D-Galactoside alpha 2-6-Sialyltransferase