Journal article

Detection of gangliosides by direct binding of Limax flavus agglutinin to thin layer chromotograms

W Kielczyski, LC Harrison

Glycoconjugate Journal | Published : 1990


A simple and sensitive method for detecting gangliosides on TLC plates is described. Gangliosides are extracted by phase partition in chloroform/methanol, developed on TLC plates in chloroform/methanol/0.25% aqueous KCl (5/4/1 by vol) and identified by binding of125I-labelled, sialic acid-specific Limax flavus agglutinin (LFA) autoradiography and scanning densitometry. The detection limit of the method is below 1 ng (0.5 pmol) for GM3, GM1 and GT1b, and below 0.3 ng (0.2 pmol) for GM2 and GD1a. Binding of125I-LFA is not inhibited by 106-fold molar excess concentrations of N-acetylneuraminic acid or lactose but is decreased in a dose-dependent manner by either N-acetylneuraminyllactose or unl..

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