The molecular properties of the major, high-frequency antigens (Ge2 and Ge3) of the human Gerbich blood group system were investigated using 14 different alloantibodies from rare Ge: -1,-2,-3 or Ge: -1,-2,3 individuals. Various modification, fractionation or fragmentation products of glycophorins (sialoglycoproteins) from normal erythrocytes (phenotype Ge: 1,2,3) were used in hemagglutination inhibition assays. The location of the antigens was also studied by blotting of proteins, separated by dodecyl sulfate polyacrylamide gel electrophoresis, to nitrocellulose and detection of bound antibodies by 125I-labelled protein G. Anti-Ge3 was found to be directed against a region of glycophorin C that surrounds a tryptic cleavage site at position 48 and a similar region of glycophorin D whose structure is not yet known. NeuAc residue(s), probably representing part(s) of a carbohydrate unit attached to serine42 of glycophorin C, methionine, aspartic or glutamic acid, tryptophan and/or arginine residue(s) are involved in the Ge3 epitopes, as judged from chemical modification. The Ge2 epitopes were found to be located on a tryptic glycopeptide from glycophorin D comprising about 20-30 amino-acid residues. NeuAc residue(s), attached to serine-/threonine-linked oligosaccharide(s), are involved in the Ge2 determinants. Using the immunoblotting technique, it could also be shown that the 'new' glycophorin in Ge: -1,-2,3 cells carries the Ge3 antigen.