Sensitive enzyme-immunostaining and densitometric determination on thin-layer chromatography of N-glycolylneuraminic acid-containing glycosphingolipids, Hanganutziu-Deicher antigens.

Abstract:

:A sensitive enzyme-immunochemical staining method was developed for detection of N-glycolylneuraminic acid (NeuGc)-containing glycosphingolipids (GSLs) on silica gel thin-layer chromatography. The procedure consists of immune reaction among NeuGc-containing GSLs, affinity-purified chicken anti-NeuGc-LacCer and horseradish peroxidase-conjugated rabbit anti-chicken IgG, and the peroxidase reaction using 4-chloro-1-naphthol as a chromogenic substrate. Quantitative determination was achieved by direct densitometric scanning of the enzyme-immunostained spots on the chromatogram. As little as 0.5 pmol of NeuGc-LacCer, NeuGc-nLcOse4Cer, and NeuGc-nLcOse6Cer (0.64-1.0 ng) could be detected with a good signal-to-noise ratio. A semi-linear detector response was observed up to 50 pmol of each GSL. This procedure can be applied easily to other glycolipid antigen systems.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Higashi H,Fukui Y,Ueda S,Kato S,Hirabayashi Y,Matsumoto M,Naiki M

doi

10.1093/oxfordjournals.jbchem.a134760

subject

Has Abstract

pub_date

1984-05-01 00:00:00

pages

1517-20

issue

5

eissn

0021-924X

issn

1756-2651

journal_volume

95

pub_type

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