This in vitro study evaluated the effect of Er:YAG laser on adhesion to human dentin of Grossman, Endomethasone, N-Rickert, and Sealer 26 root canal sealers. The crowns of 40 human molars were cut on the occlusal side until a flat dentin surface was obtained. The teeth were divided into two groups: group 1, no laser application; and group 2, irradiation with Er:YAG laser (KaVo Key Laser 2; 11 mm focal distance, perpendicular to the dentin surface, 4 Hz frequency, 200 mJ energy, 62 J total energy and 313 pulses, 1-min application time, and 2.25 W power). Five samples were tested for each sealer and each group. An Instron universal testing machine was used for the adhesion test. Sealer 26 showed the best adhesion both with and without laser application (p < 0.01). Grossman and N-Rickert sealers had intermediate values, and Endomethasone had the worst adhesion. Application of Er:YAG laser did not alter the adhesion of Grossman, N-Rickert, or Endomethasone sealers. However, laser application increased the adhesion of Sealer 26. The epoxy resin-based root canal sealer (Sealer 26) adhered better to dentin prepared with and without Er:YAG laser than the zinc oxide/eugenol-based sealers (Endomethasone, N-Rickert, and Grossman).