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Estradiol (E2) binding capacity to plasma protein was estimated by simplified methods. Three techniques of dextran coated charcoal (D.C.), ammonium sulfate (A.S.) and concanavalin A sepharose (Con A.) were compared to separate bound and unbound hormones. The D.C. and A.S. methods proceeded as follows: Purified 3H-E2 in benzene-methanol was evaporated under nitrogen gas, and a half ml of sample plasma was added. Then, 0.4 ml of D.C. or A.S. was added to the plasma after one hour incubation at 37°C. Separation of protein bound E2 was performed by centrifugation of 3000 r.p.m. at 4°C for 30 minutes. The supernate was counted by a liquid scintillation gamma counter. The following procedure was performed in the Con A. method: Sample plasma of 0.1 ml and Con A. of 0.3 ml were mixed for one hour. Then, 3 H-E2 was added and mixed for 30 minutes and settled for two hours. The supernate and/or sediment were counted. This procedure could be performed at room temperature. Reproducible results were obtained when the prepared concentration of D.C. was 0.5%, A.S. was 50% and Con A. was 50%. The highest binding capacity was observed in alpha globulin fraction compared with beta and gamma globulin. The observed binding

The Method for the Measurement of the Binding Capacity of Testosterone-Estradiol Binding Globulin