The in vivo uptake of 3H-estradiol-17beta by the various lymphoid tissues, fat tissue, skeletal muscle and circulating blood of castrated male and female mice of C57BL strain was studied after an intravenous injection of the hormone. In both sexes the highest uptake was by the bone marrow and fat tissue, followed by the spleen, thymus and mesenteric lymph node. The lowest was by the muscle and the blood serum. The lymphoid tissues in the female took up a slightly more amount of radioactivity than those in the male until 1 hr after the injection, but thereafter the uptake was at almost the same level between both sexes. The lymphoid tissues showed a relative and long-term retention of the radioactivity, as compared with that in the muscle and blood serum. The binding of 3H-estradiol-17beta by the lymphoid tissue cytosol and their binding specificity for the hormone were examined in vitro by Sephadex G-100 column analysis. All the tissue cytosols tested contained estradiol-binding component(-s). Among those the thymic cytosol showed to contain the component(-s) which is fairly specific to this hormone. These results suggest that there is a relatively high uptake and retention of estrogen in the lymphoid tissues in which it is bound to cytoplasmic binding component(-s).