This paper reports the production of a rat monoclonal antibody, 14D10, which recognizes a determinant shared between B cells and Lyt-2+ T cells. Three subpopulations of spleen cells were defined based on the density of bound 14D10. They have been designated 14D10-SP (strongly positive), 14D10-WP (weakly positive), and 14D10-N (negative). Dual parameter immunofluorescence studies showed that 90% of 14D10-SP cells are IgM+ cells and more than 92% IgM+ cells are 14D10-SP. Of 14D10-WP cells, approximately 60% are Lyt-1+2+ T cells and less than 10% are IgM+ cells. However, most (greater than 90%) of Lyt-2+ T cells are reactive with 14D10. Staining of purified populations of Lyt-2+ and Lyt-2- T cells with fluoresceinated 14D10 also showed greater than 90% of Lyt-2+ T cells to be 14D10+, and fewer than 10% of Lyt-2- T cells are 14D10+. Mouse thymus contained very few (congruent to 5%) 14D10+ cells. Analysis of bone marrow cells shows almost all small cells to be 14D10+. Six Abelson virus-transformed cell lines all express the antigen recognized by 14D10.