caa a22 4500 465734200 CHVBK 20180323111750.0 cr unu---uuuuu 170327e19900101xx s 000 0 eng 10.1007/BF01050404 doi (NATIONALLICENCE)springer-10.1007/BF01050404 Detection of gangliosides by direct binding of Limax flavus agglutinin to thin layer chromotograms [Elektronische Daten] [Wojciech Kielczyski, Leonard Harrison] 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-specificLimax 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 ofN-acetylneuraminic acid or lactose but is decreased in a dose-dependent manner by eitherN-acetylneuraminyllactose or unlabelled lectin. Gangliosides were not detected after their treatment byClostridium perfringens sialidase in the presence of taurocholic acid. Ten gangliosides were detected in human brain and seven in normal human serum. Extracts from 0.2 mg of brain and 20 μl of serum were sufficient for the detection of major gangliosides. Glycoconjugate Journal, 1990 gangliosides nationallicence Limax flavus lectin nationallicence sialic acid nationallicence thin layer chromatography nationallicence autoradiography nationallicence LFA : Limax flavus agglutinin nationallicence ELLA : Enzyme Linked Lectin Assay nationallicence PIM : Poly(isobutyl methacrylate) nationallicence PVP : Polyvinylpyrrolidone mol.wt. 40,000 nationallicence PBS : Phosphate buffered saline nationallicence BSA : Bovine serum albumin nationallicence Kielczyski Wojciech Burnet Clinical Research Unit, The Walter and Eliza Hall Institute of Medical Research, P.O. The Royal Melbourne Hospital, 3050, Melbourne, Victoria, Australia aut Harrison Leonard Burnet Clinical Research Unit, The Walter and Eliza Hall Institute of Medical Research, P.O. The Royal Melbourne Hospital, 3050, Melbourne, Victoria, Australia aut Glycoconjugate Journal Kluwer Academic Publishers 7/1(1990-01-01), 75-84 0282-0080 7:1<75 1990 7 10719 https://doi.org/10.1007/BF01050404 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF01050404 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kielczyski Wojciech Burnet Clinical Research Unit, The Walter and Eliza Hall Institute of Medical Research, P.O. The Royal Melbourne Hospital, 3050, Melbourne, Victoria, Australia aut NATIONALLICENCE 700 1- Harrison Leonard Burnet Clinical Research Unit, The Walter and Eliza Hall Institute of Medical Research, P.O. The Royal Melbourne Hospital, 3050, Melbourne, Victoria, Australia aut NATIONALLICENCE 773 0- Glycoconjugate Journal Kluwer Academic Publishers 7/1(1990-01-01), 75-84 0282-0080 7:1<75 1990 7 10719 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer