2. Academic Validation
  3. Detection of Sialic Acids on the Cell Surface Using Flow Cytometry

Detection of Sialic Acids on the Cell Surface Using Flow Cytometry

  • Methods Mol Biol. 2022:2556:31-35. doi: 10.1007/978-1-0716-2635-1_3.
Hirohito Ishigaki 1,  Yasushi Itoh 2
Affiliations

Affiliations

  • 1 Division of Pathogenesis and Disease Regulation, Department of Pathology, Shiga University of Medical Science, Shiga, Japan. ihiro@belle.shiga-med.ac.jp.
  • 2 Division of Pathogenesis and Disease Regulation, Department of Pathology, Shiga University of Medical Science, Shiga, Japan.
PMID: 36175624 DOI: 10.1007/978-1-0716-2635-1_3
Abstract

We described a method to detect α2-3 linked and α2-6 linked sialic acids on the cell surface with using flow cytometry. Cells were fixed with 4% paraformaldehyde, and then α2-3 and α2-6 sialic acids were stained with biotinylated MAACKIA AMURENSIS LECTIN II (MALII) and biotinylated ELDERBERRY BARK LECTIN (SNA), respectively. Sialic acids on the cell surface were cleaved by sialidase in acetate buffer at pH 5.5 to confirm the specificity of staining. Streptavidin conjugated with Alexa flour 488 was used to detect biotinylated lectins. Thus, the α2-3 linked and α2-6 linked sialic acids on the cell surface were semi-quantitatively detected by flow cytometry.

Keywords

MALII, SNA; Sialic acid; flow cytometry; α2-3 linkage; α2-6 linkage.

Figures
Products