Neoglycolipid-based oligosaccharide microarray system: preparation of NGLs and their noncovalent immobilization on nitrocellulose-coated glass slides for microarray analyses

Methods Mol Biol. 2012:808:117-36. doi: 10.1007/978-1-61779-373-8_8.

Abstract

Carbohydrate microarrays, since their advent in 2002, are revolutionizing studies of the molecular basis of protein-carbohydrate interactions both in endogenous recognition systems and pathogen-host interactions. We have developed a unique carbohydrate microarray system based on the neoglycolipid (NGL) technology, a well-validated microscale approach for generating lipid-tagged oligosaccharide probes for use in carbohydrate recognition studies. This chapter provides an overview of the principles and key features of the NGL-based oligosaccharide microarrays, and describes in detail the basic techniques - from the preparation of NGL probes to the generation of microarrays using robotic arraying hardware, as well as a general protocol for probing the microarrays with carbohydrate-binding proteins.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Collodion / chemistry*
  • Glass / chemistry*
  • Glycolipids / chemical synthesis
  • Glycolipids / chemistry*
  • Oligonucleotide Array Sequence Analysis / methods*
  • Oligosaccharides / chemical synthesis
  • Oligosaccharides / chemistry*

Substances

  • Glycolipids
  • Oligosaccharides
  • Collodion