The human uterine progesterone receptor was subjected to high-performance liquid chromatography on size exclusion, anion exchange and chromatofocusing columns. For the rapid isolation of the receptor, recovery of [3H]progesterone as well as protein from the columns was essential. The size exclusion columns (G2000 SW and G3000 SW) as well as Mono P HR 5/20 chromatofocusing column adsorbed [3H]progesterone and thus were not useful for separation purposes. The anion exchange (polyanion SI-17) and chromatofocusing columns, AX500, and IEX 540 DEAE gave very good recoveries of protein (greater than 90%) and [3H]progesterone; 80, 66 and 88% respectively. These columns gave rapid and reproducible separation of the progesterone receptor from other cytosol proteins.