Determination of Glycolipid Ligands of Carbohydrate-Binding Proteins Directly on Thin Layer Chromatograms

Methods Mol Biol. 2018:1804:419-427. doi: 10.1007/978-1-4939-8552-4_21.

Abstract

Determination of a glycolipid ligand can quickly reveal the carbohydrate epitope recognized by carbohydrate-binding proteins (CBP) which may include antibodies, lectins, toxins, and virulence factors. Thin layer chromatography (TLC) is a simple, well-known, and heavily used method to separate glycolipids based on their migration through a thin layer of chromatography media, usually silica gel, coated on the surface of a rigid plate. Reacting carbohydrate-binding proteins directly on a TLC plate of separated glycolipids is a powerful technique to quickly determine functional carbohydrate epitopes present in an entire glycolipid extract of a target cell type or tissue.

Keywords: Anti-carbohydrate antibodies; Carbohydrate-binding proteins (CBP); Glycolipid antigens; Immunostaining; Lectins; Thin layer chromatography (TLC); Toxins.

MeSH terms

  • CA-19-9 Antigen / analysis
  • Chromatography, Thin Layer / methods*
  • Colorimetry
  • Glycolipids / analysis*
  • Humans
  • Ligands
  • Peroxidase / metabolism
  • Receptors, Cell Surface / chemistry*
  • Reference Standards
  • Staining and Labeling
  • Substrate Specificity

Substances

  • CA-19-9 Antigen
  • Glycolipids
  • Ligands
  • Receptors, Cell Surface
  • saccharide-binding proteins
  • Peroxidase