The intent of this project is to purify and characterize glucocorticoid receptors by using affinity chromatography, conventional protein purification methods, generation of monoclonal antibodies to be used in antibody affinity chromatography, and site specific covalent steroid affinity labelling of the receptor. We also seek to establish in vitro systems, derived from rat kidney and liver, in which to coordinately measure receptor occupancy and effects. Ultimately these systems can be used for reconstruction experiments with purified receptor. During the coming year we intend to continue production of partially purified glucocorticoid receptor by two-step DNA cellulose chromatography. This material will be used for; 1. generation of monoclonal antibodies to the receptor, 2. characterization and stabilization of the partially purified receptor, and 3. purification of the receptor to homogeneity by additional steps possibly including isoelectrofocussing or column chromatography. A comparison will also be made between BudR substituted and non-substituted DNA cellulose in terms of efficiency of purification of the glucocorticoid receptor.