The steroid-protein binding equilibrium in extra-cellular fluids is considered to have possible relevance to the development of endometrial and breast cancer. A method is described to measure the fraction of steroid bound to sex hormone binding globulin (SHBG) in serum, using binding of SHBG to Con-A-sepharose. To validate the assay, serum samples with different SHBG levels were studied at various dilutions and with various amounts of added ligands. Sera with high SHBG concentrations still showed considerable steroid binding after heating to 59 degrees C for 30 min. The intra-assay variation of the assay ranged from 2-4%, the inter-assay variation was approx. 5%.