SUMMARY The binding characteristics of corticosterone by rat liver were studied by a displaceable binding technique. The binding of corticosterone to protein fractionated by gel filtration and density gradient centrifugation has been carried out as a preliminary determination of the nature of the binding sites. The results were analysed and showed three types of binding sites for corticosterone with the characteristic association constants at 0° of K1 = 1·2 × 1010, K2 = 1 × 108 and K3 = 1 × 104 1./mole. Percentage displacement of corticosterone from the nuclear fraction did not differ significantly from that from tissue or the mitochondrial-microsomal fraction. The K1 and K2 sites persisted in separated buffer-soluble fractions but were destroyed on mild heating leaving only the K3 sites.