Monoclonal H-Y antibody of demonstrated specificity was reacted with soluble H-Y antigen in a newly-developed enzyme-linked immunosorbent assay (ELISA). Typing of cell lines was accomplished by qualitative and quantitative absorption. In one case, the antibody was subdivided into equal portions, each of which was absorbed with 1, 3, 6 or 12 X 10(6) cells from male or female; after absorption, the cells were discarded and the antibody was titrated for residual activity in the ELISA. With increasing cell numbers, optical density scores for male-absorbed antibody approached baseline levels. Although male-female differences were pronounced and highly significant, there was also a fall in reactivity after absorption with female cells.