The ability of antiganglioside antibodies to detect their respective antigens in various environments was studied. In contrast to sensitive detection of pure ganglioside standards by ELISA, antibody binding to mixtures of gangliosides was drastically reduced. This loss of sensitivity also occurred with immunostaining of gangliosides absorbed to silica gel. Moreover, absorption of antibodies to antigen-containing lipid vesicles failed, if the vesicles were prepared together with other gangliosides. These data indicate that antigen accessibility is strongly influenced by surrounding gangliosides. This should be considered whenever gangliosides are traced by antibodies. The ELISA procedure appears useful to test for such properties.