In situ visualization of glycosphingolipids on thin-layer chromatograms using lectin-peroxidase conjugates

A technique is described for direct detection by lectins conjugated to horseradish peroxidase of the oligosaccharides of glycosphingolipids on thin-layer chromatograms. Final visualization is accomplished using a substrate medium consisting of hydrogen peroxide and 3,3'-diaminobenzidine. The procedure is rapid, specific, sensitive and highly reproducible, and staining patterns are stable for years. In addition to providing preliminary structural information, this technique can be employed in combination with other methods, such as autoradiographic detection of tritium-labeled glycosphingolipids in the same chromatogram.