This study sought to compare the antibacterial efficacy of three commonly used intracanal medicaments with propolis against Enterococcus faecalis. This study utilized 180 freshly extracted single-rooted intact human permanent teeth with a single root canal. After root canal preparations and sterilization, canals were contaminated with E. faecalis and incubated at 37 degrees C (+/- 1.0 degrees C) for seven days. The teeth were divided randomly into six groups. To determine bacterial growth on blood agar, microbiological samples were carried out with sterile paper points to evaluate results at 48 hours and at ten days. All data were analyzed statistically with t-test, Mann Whitney, Kruskal Wallis, and one-way ANOVA tests. This study revealed that propolis had good in vitro antibacterial activity against E. faecalis in the root canals, suggesting that it could be used as an alternative intracanal medicament.