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High-efficiency labeling of sialylated glycoproteins on living cells

Academic Article
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Overview

authors

  • Zeng, Y.
  • Ramya, T. N. C.
  • Dirksen, A.
  • Dawson, Philip
  • Paulson, James

publication date

  • 2009

journal

  • Nature Methods  Journal

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.

subject areas

  • Aldehydes
  • Aniline Compounds
  • Animals
  • Antigens, CD45
  • Biotin
  • Biotinylation
  • CHO Cells
  • Catalysis
  • Cell Line
  • Cell Survival
  • Cricetinae
  • Cricetulus
  • Endocytosis
  • Flow Cytometry
  • Humans
  • Membrane Glycoproteins
  • Microscopy, Fluorescence
  • N-Acetylneuraminic Acid
  • Oxidation-Reduction
  • Oximes
  • Periodic Acid
  • Sialyltransferases
  • Staining and Labeling
  • Streptavidin
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Identity

PubMed Central ID

  • PMC2830088

International Standard Serial Number (ISSN)

  • 1548-7091

Digital Object Identifier (DOI)

  • 10.1038/nmeth.1305

PubMed ID

  • 19234450
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Additional Document Info

start page

  • 207

end page

  • 209

volume

  • 6

issue

  • 3

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