Glycosphingolipid-blotting: an immunological detection procedure after separation by thin layer chromatography.
A method for detecting glycosphingolipids (GSL) in situ after thin layer chromatography is described. The separated GSL are transferred by diffusion to nitrocellulose. The replica is incubated with poly- or monoclonal antibodies and bound antibodies are detected with second antibodies coupled to peroxidase. Advantages of the procedure are its speed, the non-radioactive detection method, and its suitability for screening applications. In addition, small scale affinity purification of antibodies from the replicas is possible. The presence of Forssman antigen in mouse tissues and the reaction of monoclonal antibodies with human GSL is demonstrated.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000937:Antigen-Antibody Reactions; D001667:Binding, Competitive; D002241:Carbohydrates / Q000494:pharmacology; D002855:Chromatography, Thin Layer; D003101:Collodion; D005577:Forssman Antigen / Q000276:immunology; D006028:Glycosphingolipids / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D006801:Humans; D007124:Immunoenzyme Techniques; D051379:Mice; D011817:Rabbits
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D006028:Glycosphingolipids; D003101:Collodion; D005577:Forssman Antigen