Pancreatic glucagon receptor radioassay has been developed with rat liver cell plasma membrane prepared with sucrose gradient centrifugation. 125I-glucagon was prepared in this laboratory and the separation of B/F was done by low speed centrifugation with addition of PEG (W/V, 20%) and normal human serum. The specific binding of 125I-glucagon was 25%-35% and nonspecific binding was less than 5%. Reproducibility within and between- assay was 6.83% and 8.40% (CV%), respectively. Binding characteristics of glucagon receptor of normal and streptozotocin-induced diabetic rats were determined by using two methods, receptor dilution and conventional competitive binding assay. The results showed that there were no marked differences concerning high and low binding affinities between normal and diabetic rats, but the concentration of binding sites for both high and low affinity receptors was significantly higher for diabetic rats than normal rats. The possible mechanisms for the above results have been discussed.