Previous biochemical studies have shown that B cell specific, monoclonal antibody, McAb 33.1, reacts with a class II antigen that represents a human analogue of murine I-A (DS/DC) antigens (J. Exp. Med. 158:1924, 1983). McAb 33.1 recognizes a polymorphic human B lymphocyte specific antigen present on mu+, B1+ peripheral B cells, B lymphoid cell lines, activated B cells, and neoplastic B lymphoid cells. Of 100 HLA-D/DR typed EBV-transformed lymphoblastoid cell lines tested, only those from DR3,3 and DR7,7 individuals failed to react with McAb 33.1. The 33.1 antigen is present at lower concentrations on B cells from blood, tonsil, spleen, and lymph node when compared to B cell lines. By contrast, the antigen is not detectable on blood T lymphocytes, T cell lines, or mitogen activated T cells and it is absent on monocytes of some individuals or is present only on a minor subpopulation (approximately 20%). McAb 33.1 should facilitate the functional, structure, and molecular dissection of the human Ia system.