Objectives: The aim of the present study was to investigate the bond strength of RelyX Unicem (3M) to root canal dentin when used as an endodontic sealer. Materials and Methods: Samples of 24 single-rooted teeth were prepared with Gates Glidden drills and K3 files. After that, the roots were randomly assigned to three experimental groups (n = 8) according to the filling material, (1) AH Plus (Dentsply De Trey GmbH)/Gutta-Percha cone; (2) Epiphany SE (Pentron)/Resilon cone; (3) RelyX Unicem/Gutta-Percha cone. All roots were filled using a single cone technique associated to vertical condensation. After the filling procedures, each tooth was prepared for a push-out bond strenght test by cutting 1 mm-thick root slices. Loading was performed on a universal testing machine at a speed of 0.5 mm/min. One-way analysis of variance and Tukey test for multiple comparisons were used to compare the results among the experimental groups. Results: Epiphany SE/Resilon showed significantly lower push-out bond strength than both AH Plus/Gutta-Percha and RelyX Unicem/Gutta-Percha (p < 0.05). There was no significant difference in bond strength between AH Plus/Gutta-Percha and RelyX Unicem/Gutta-Percha (p > 0.05). Conclusions: Under the present in vitro conditions, bond strength to root dentin promoted by RelyX Unicem was similar to AH Plus. Epiphany SE/Resilon resulted in lower bond strength values when compared to both materials.