In situ immunological determination of basic carbohydrate structures of gangliosides on thin-layer plates

Megumi Saito, Noriyuki Kasai, Robert K. Yu

Research output: Contribution to journalArticlepeer-review

91 Scopus citations

Abstract

An immunological method for the determination of the basic carbohydrate structure of gangliosides by using a thin-layer chromatographic immunostaining technique was developed. After high-performance thin-layer chromatography of gangliosides, the chromatogram is treated with a 0.4% polyisobutylmethacrylate solution. Arthrobacter ureafaciens neuraminidase is then applied to the separated gangliosides in situ on the chromatographic plate. This procedure will remove both external and internal sialic acid residues from the core oligosaccharide backbone. The resulting glycolipid products are then incubated with anti-Gg4 serum and 125I-staphylococcal protein A, successively, and exposed to an X-ray film. Through a highly specific binding, the anti-Gg4 antibody detects only those gangliosides having the oligosaccharide backbone of Gg4.

Original languageEnglish (US)
Pages (from-to)54-58
Number of pages5
JournalAnalytical Biochemistry
Volume148
Issue number1
DOIs
StatePublished - Jul 1985
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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