A mouse monoclonal antibody (MAb 3AE8) of the IgG1 isotype was prepared against rabbit splenocytes and was found by indirect immunofluorescence and direct binding assays to react, in the rabbit, primarily with oil-induced peritoneal exudate macrophages (PEM phi). This MAb did not bind to rabbit T cells, B cells, polymorphonuclear leukocytes, or resident alveolar or peritoneal M phi but it did bind to a subpopulation of rabbit splenocytes with surface characteristics of null cells. The antibody also recognized mouse and rat PEM phi as well as the murine M phi cell lines P388D1 and IC-21. Consistent with findings in the rabbit, it did not bind to M phi obtained from the peritoneal cavities of rats or mice. The addition of MAb 3AE8 to mouse PEM phi caused a marked enhancement in the phagocytic uptake of erythrocyte target cells sensitized with a mouse antierythrocyte antiserum.