We describe a double-antibody radioimmunoassay for antimicrosomal antibodies in serum. Antigen in centrifuged pellets of microsome is solubilized by treatment with sodium deoxycholate solution and radiolabeled (125I] by the Chloramine T method. A 10-microL aliquot of human serum or antimicrosomal antibody standard is incubated overnight at room temperature with 125I-labeled microsome. Bound radiolabeled microsome is separated by precipitation with goat antihuman IgG antiserum. The percentage of 125I-labeled microsome bound by each serum is calculated, and its concentration of antimicrosomal antibody interpolated from a standard curve. The prevalence of abnormal antimicrosomal antibody was 16% in a self-referred asymptomatic or not acutely ill group of patients having a periodic multiphasic health examination. Values for patients with certain thyroid diseases generally agreed with other published data. We conclude that our assay is sensitive, precise, and accurate, and is more efficient for routine measurement of antimicrosomal antibody than the currently used solid-phase radioimmunoassay.