Prunella vulgaris ( Labiatae family) or self-heal is traditionally used for different ailments such as eye pain and inflammation, headache, dizziness, sore throat and wound healing. Total phenolic and total flavonoid contents of extracts (methanol, ethanol and aqueous) were determined by a spectrophotometer. The antimicrobial activity was evaluated by micro broth dilution assay. The total phenolic content of P. vulgaris extracts were higher in aqueous extract (156.5 mg GAC/g) followed by ethanol extract and methanol extract. The TFC content of P. vulgaris methanol extract (82.8 mg QE/g) was higher than ethanol extract (22.7 mg QE/g) and aqueous extract (16.2 mg QE/g). The antimicrobial activity of methanol or ethanol extracts was higher than aqueous extract from P. vulgaris. The sensitivity of microorganisms to different extracts is related to type of pathogens. There is no positive relation between total phenolic content and its antimicrobial activity. Prunella vulgaris ethanolic extract as a source of phenolic and flavonoid contents can be used as an antimicrobial agent.