Autoradiography of Ganglioside Antigens Separated by High-Performance Thin-Layer Chromatography with Their Antibodies

Abstract

An improved method is described for the detection of ganglioside antigens with the antibodies on thin-layer chromatograms. The method involves the following steps: Separation of gangliosides on a high-performance thin-layer chromatographic plate, application of antibody solution, and detection of bound antibody with [¹²⁵I] staphylococcal protein A. Using specific antibodies against gangliosides GM 4, GM 1, GD 3, and asialo GM 1 ganglioside, the method allowed positive identification of these antigens on thin-layer plates. It also provides a convenient means of assessing the specificity of an anti-glycolipid antibody.