Bauhinia species are trees up to 6-12m in height. Traditionally, leaves and stem barks have been used against pain, inflammation, dysentery, mycosis, diarrhea and diabetes. Bauhinia rufescens Lam. is a tropical forage up to 8 m in height. In African system of medicine, the plant has been used against gout, leprosy, and malaria. This study was designed to investigate the major flavonoid of Bauhinia rufescens roots and to assess the antimicrobial activity of the crude ethanolic extract and the isolated flavonoid. The flavonoids were extracted with ethanol and the crude extract was purified by paper chromatography where a pure flavonoid (compound I) was isolated. The structure of this compound has been partially characterized by some spectral tools (UV, IR and ¹HNMR). In the antimicrobial assay, the crude ethanolic extract showed better inhibitory effect against a panel of human pathogens compared to compound I. While the crude extract showed significant antibacterial and antifungal properties, compound I exhibited moderate antibacterial activity and weak antifungal properties.