The surface antigen phenotype of the immediate precursors of clonable B lymphocytes was investigated with conventional alloantisera and monoclonal antibodies directed by B lineage antigens. Ia was demonstrable on B cells, but not their immediate precursors in adult marrow. Adult, but not fetal, B cell precursors were susceptible to lysis with anti-Lyb-2 or anti-Qa. A panel of monoclonal rat antibodies was prepared and placed into categories on the basis of recognition patterns obtained with established cell lines. Of 2 groups that are described here, 1 (typified by antibodies from clone 14.8) detect an antigen that is preferentially expressed on B cells and their precursors, a proportion of antibody-secreting cells, and a subpopulation of peripheral T lymphocytes. Cells that did not display demonstrable amounts of antigen include brain, granulocytes, macrophages, mastocytoma cells, and erythroleukemia cells. A 2nd category of antibodies revealed an antigen that was more widely distributed on hemopoietic cells. Cells capable of quickly maturing into functional, colony-forming B lymphocytes in culture or after transfer to irradiated recipients specifically adhered to 14.8 antibody-coated, polystyrene petri dishes in the cold. Reductions in numbers of stem cells (CFU-s) and myeloid progenitors (CFU-c) by this treatment were minimal. Of particular importance was the fact that these antibodies recognized cells in embryonic liver as well as in adults that were destined to become B lymphocytes. These observations provide new perspective on B lineage precursor heterogeneity and suggest ways of localizing and dissecting some of the earliest events that are critical to development of the humoral immune system.