The Extraction and Analysis of Glycosphingolipids

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.