Since the prevailing form of hormone replacement therapy is associated with the development of cancer in breast and endometrial tissues, alternatives are needed for the management of menopausal symptoms. Formulations of Trifolium pratense L. (red clover) are being used to alleviate menopauseassociated hot flashes but have shown mixed results in clinical trials. The strobiles of Humulus lupulus L. (hops) have been reported to contain the prenylflavanone, 8-prenylnaringenin (8-PN) as the most estrogenic constituent, and this was confirmed using an estrogen receptor ligand screening assay utilizing ultrafiltration mass spectrometry. Extracts of hops and red clover and their individual constituents including 8-PN, 6-prenylnaringenin (6-PN), isoxanthohumol (IX), and xanthohumol (XN) from hops, and daidzein, formononetin, biochanin A, and genistein from red clover, were compared using a variety of in vitro estrogenic assays. The IC 50 values for the estrogen receptor α and β binding assays were 15 and 27 μg/mL, respectively, for hops and 18.0 and 2.0 μg/mL, respectively, for the red clover extract. Both of the extracts, genistein, and 8-PN activated the estrogen response element (ERE) in Ishikawa cells while the extracts, biochanin A, genistein, and 8-PN significantly induced ERE-luciferase expression in MCF-7 cells. Hop and red clover extracts, as well as 8-PN upregulated progesterone receptor (PR) mRNA in the Ishikawa cell line. In the MCF-7 cell line, PR mRNA was significantly upregulated by the extracts, biochanin A, genistein, 8-PN, and IX. The two extracts had EC 50 values of 1.1 and 1.9 μg/mL, respectively, in the alkaline phosphatase induction assay. Based on these data, hops and red clover could be attractive for development as herbal dietary supplements to alleviate menopause-associated symptoms.