Background/purpose: Bacterial infection is closely associated with the failure of endodontic treatment, and use of endodontic sealer with antimicrobial activity and biological compatibility is necessary for the success of root canal treatment. The purpose of this study was to investigate and to compare the antibacterial effect of two calcium silicate-based root canal sealers (Endoseal and EndoSequence BC sealer) as recent development sealers and with three conventional root canal sealers (AH Plus, Sealapex, and Tubli-Seal), before or after setting, on Porphyromonas endodontalis, Porphyromonas gingivalis, and Enterococcus faecalis. Materials and methods: The sealers were soaked in phosphate buffered saline to elute its compositions after and before setting, and the elutes were performed the antimicrobial assay. Also, X-ray fluorescence analysis was carried out to compare compositions of two calcium silicate-based sealers. Results: The conventional root canal sealers have strong antibacterial activity against the Gram-negative bacteria, P. endodontalis and P. gingivalis. Endoseal sealer showed antibacterial activity against not only the Gram-negative bacteria, but also against the Gram-positive bacteria, E. faecalis. However, Endosequence BC sealer exhibited a weak antibacterial effect on all bacteria in this study. X-ray fluorescence analysis exhibited that Endoseal contained more types and more amount of the oxide compound known to have strong antimicrobial activity such as Al2O3, Fe2O3, MgO, Na2O, NiO, and SO2 than Endoseqeunce BC. Conclusion: Endoseal, which contains various types of oxide compounds, seems to be a suitable sealer for preventing bacterial infection in both treated and untreated root canals. Keywords: Root canal sealer, Antimicrobial activity, Oxide compound, E. faecalis