Stimulation of lymphocytes with a mitogenic lectin such as concanavalin A (ConA) results in differentiation and cell division. Among the changes which occur after stimulation are increases in phosphorylation of proteins and in protein kinase activity. We used a high-resolution, nondenaturing gel system to separate and visualize protein kinases in situ. We have clearly identified both autophosphorylating and substrate-dependent kinases. One band of cyclic AMP-dependent kinase activity was significantly enhanced in lectin-stimulated cells. In contrast, treatment of the cells with phorbol ester under conditions which depress stimulation caused a decrease in the activity of one kinase.