The kinetics of clearance of 125I from the popliteal lymph nodes and feet of human serum albumin (HSA)-immunized mice was studied following the injection of [125I]-HSA into the hind footpads. Antigen was cleared from both locations rapidly for the first few days. The antigen half-life (T½) during this period was only a matter of hours. By the end of the first week, however, the rate of clearance in both sites had changed markedly. The antigen T½ in the node between the first and sixth week was 8.1 weeks (95% confidence interval between 5.1 and 20 weeks) and the antigen T½ in the foot was 6.1 weeks (95% confidence interval between 3.7 and 16.6 weeks). There was, however, about twenty times more radioactivity in the feet than in the popliteal nodes. Autoradiography of popliteal lymph nodes revealed that initially antigen was trapped in the medulla, subcapsular sinus, superficial cortex and around lymphoid follicles. During the first few days antigen was cleared from all sites except the follicles. The radioactivity initially trapped in the medulla, subcapsular sinus, and superficial cortex appeared to have been associated with macrophages. Studies with peritoneal macrophages indicated an antigen T½ in these cells of 2 h (95% confidence interval between 1.5 and 3 h). The initial rapid clearance of antigen trapped and catabolized by macrophages and the long-term retention of antigen in the follicles is probably attributable to trapping and retention by follicular dendritic cells. The large pool of antigen trapped in the foot did not appear to serve as a depot to replace antigen degraded in the node, since amputation of the foot did not alter the level of antigen retained in the node. The long antigen T½ in the lymph node follicles indicates that antigen is available in the lymph node to play a role in the maintenance and regulation of immune responses for many months or even years.