Purpose: This study aimed to compare between Calcium hydroxide, Propolis and Triple Antibiotic Paste as root canal medicaments in eradication of Enterococcus faecalis. Material and Methods: for this study, 45 extracted human teeth were used. After their decapitation, they were initially instrumented to reach 20 K-file, later they were separated into 3 sets according to the medicament used; Group 1: Triple antibiotic paste, Group 2: Propolis paste, Group 3: Calcium hydroxide paste (15 specimens in each group). After sterilization of all samples inoculation was performed with Enterococcus faecalis (ATCC29212) over a period of 21 days. Preparation of all samples was done using protaper files till F2 size, irrigation using sodium hypochlorite solution of 2.5% concentration and 17% EDTA, then intracanal medicaments were applied. First sample was collected after inoculation of bacteria into root canals. Second sample was collected after applicated medicament is rinsed. Muller Hilton Yeast agar was used for culturing of samples then went under incubation to count CFU’s. Results: There was a significant difference between different samples where (p ≤ 0.05) and there was a continuous decrease of (mean ±SD) value. Pairwise comparisons for all three medicaments showed all samples to be significantly different from each other. Conclusion: Antimicrobial activity of Propolis against E. faecalis was found to be greater than that of TAP and Calcium hydroxide.