Abstract
Glycolipids are an important group of diverse molecules present in the plasma membrane and to a more limited extent in the intracellular membranes of the Golgi and lysosomal systems of eukaryotic cells. They are prominent as antigenic determinants and as the ligands for specific lectin binding, and there are many examples in which they have been used as cell specific markers. The cellular glycolipid pattern frequently shows changes with the state of differentiation and may be altered following viral transformation. In animal cells the most common and most varied glycolipids are glycosphingolipids, and the methods of isolation and analysis given are mainly for this class of molecule. Methods are given for the extraction of total glycolipid from small volumes of tissue and for the isolation of the glycolipids from such extracts. Methods of fractionating and quantitating the acidic and neutral glycolipids are described, which do not require the use of specialized equipment such as high-performance liquid chromatography (HPLC), gas chromatography (GC), or mass spectroscopy and which can be performed easily in the general laboratory.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Lauter, C. J. and Trams, E. G. (1962) A spectrophotometric determination of sphingosine. J. Lipid Res. 3, 136–138.
Jourdian, G. W., Dean, L., and Roseman, S. (1971) The sialic acids; IX a periodate-resorcinol method for the quantitative estimation of free sialic acids and their glycosides. J. Biol. Chem. 246,430–435.
Kean, E. L. (1968) Rapid, sensitive spectrophotometric method for quantitative determination of sulphatides. J. Lipid Res. 9, 319–327.
Saito, T. and Hakomori, S. (1971) Quantitative isolation of total glycosphingolipids from animal cells. J. Lipid Res. 12, 257–259.
Kannagi, R., Watanabe K., and Hakomori S. (1987) Isolation and purification of glycosphingolipids by high-performance liquid chromatography, in Methods in Enzymology, vol. 138 (Ginsburg, V., ed.), Academic, New York, pp. 3–12.
Magnani, J. L. (1987) Mouse and rat monoclonal antibodies directed against carbohydrates, in Methods in Enzymology, vol. 138 (Ginsburg, V., ed.), Academic, New York, pp. 484–491.
Spitalnik, S. L. (1987) Human monoclonal antibodies directed against carbohydrates, in Methods in Enzymology, vol. 138 (Ginsburg, V., ed.), Academic, New York, pp. 492–503.
Saito, M., Kasai, N., and Yu, R. K. (1985) In situ determination of basic carbohydrate structures of gangliosides on thin-layer plates. Anal. Biochem. 148, 54–58.
Childs, R. A., Drickamer, K., Kawasaki, T., Thiel, S., Mizuochi, T., and Fiezi, T. (1989) Neoglycolipids as probes of oligosaccharide recognition by recombinant and natural mannose-binding proteins of the rat and man. Biochem. J. 262, 131–138.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1993 Humana Press Inc.
About this protocol
Cite this protocol
Gregson, N.A. (1993). The Extraction and Analysis of Glycosphingolipids. In: Graham, J.M., Higgins, J.A. (eds) Biomembrane Protocols. Methods in Molecular Biology, vol 19. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-236-1:287
Download citation
DOI: https://doi.org/10.1385/0-89603-236-1:287
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-0-89603-236-1
Online ISBN: 978-1-59259-506-8
eBook Packages: Springer Protocols