The study aims to evaluate the wound healing properties of bioactive flavonoid fraction from the ethanolic extract of Butea monosperma (Lam) stem bark. In this study the wound healing activity of the flavonoid fraction isolated from the stem bark of Butea monosperma were evaluated in excision, incision and dead space wound healing models using Albino wistar rats. The wound healing activity was assessed by the breaking strength in case of incision wounds, epithelialization and wound contraction in case of excision wound and granulation tissue dry weight, breaking strength and hydroxyproline content in case of dead space wound. The flavonoid fraction showed the significant wound healing activity on all three wound models. The phytochemical investigations revealed that the flavonoid fraction contains two isoflavones genistein and prunetine. The increased rate of wound contraction and hydroxyproline content in the flavonoid fraction treated animals provides a scientific base to the ethno medicinal use of Butea monosperma, which is largely attributable to the additive or synergistic effect of isoflavones present in the flavonoid fraction.