Withania somnifera an Indian traditional medicinal herb, is known to possess antistress, antioxidant, analgesic, anti-inflammatory, cardioprotective, adaptogenic, antispasmodic, immunomodulatory and immunostimulant activities. Its other activities include anti-inflammatory, immunoregulatory, anti-tumor, anti-angiogenic, antiinvasive and chemopreventive effects. We investigated the antioxidant, anti-inflammatory and anti-microbial activities of hydro-alcoholic extract of roots of Withania somnifera and tried to relate the role of antioxidant activity in promoting anti-inflammatory activity. 1,1-Diphenyl-2-picrylhydrazyl and nitric oxide radical scavenging assays were used to evaluate antioxidant potential whereas anti-inflammatory activity was assessed by HRBC membrane stabilization method and albumin denaturation assay. Antibacterial activity against test organisms E. coli and S. aurens was evaluated and for antifungal activity, extract was tested against Aspergillus fumigatus. Our results suggest that WS extract possesses potent antioxidant activity, significant anti-inflammatory activity and noteworthy anti-microbial activity against E. coli and S. aureous. Unfortunately, the extract was found to be inactive against Aspergillus fumigatus.