The reactivity of a panel of antiganglioside monoclonal antibodies with a number of melanoma cell lines having different ganglioside composition profiles was studied. One cell line synthesized only GM3, one produced both GM3 and GD2, 2 had GM3 and GD3 as their major gangliosides, and 2 others synthesized approximately equal amounts of GM3, GM2, GD3, and GD2 gangliosides. Antibody reactivity with viable cells was analyzed by: (a) flow cytometry on suspension cells; and (b) mixed hemagglutination assays or immune adherence assays on monolayer cells in culture. GM3 was efficiently detected only in the cell line having GM3 as its sole ganglioside. In the other cell lines, GM3 was difficult to detect even in cells in which it made up a high proportion (up to 50%) of the total ganglioside content. GM2 was easily detectable only in JB-RH melanoma cells (which contain only GM3 and GM2). GD3 was the most reactive ganglioside in 2 cell lines and GD2 in 2 other lines. In general, the most complex ganglioside present in a cell was the one most accessible to antibody. The differential exposure at the cell surface of specific gangliosides may have implications for antibody-directed tumor detection and therapy and for cell-protein or cell-cell interactions that involve glycolipids.