Er-YAG laser irradiation has been claimed to improve the adhesive properties of dentin. We tested the hypothesis that dentin adhesion is affected by Er-YAG laser conditioning. Superficial or deep dentin from human molars was: (a) acid-etched with 35% H3PO4; (b) irradiated with an Er-YAG laser (KaVo) at 2 Hz and 180 mJ, with water-cooling; and (c) laser- and acid-etched. Single Bond (3M ESPE) and Z100 composite (3M ESPE) were bonded to the prepared surfaces. After storage, specimens were tested in shear to failure. Bonded interfaces were demineralized in EDTA and processed for transmission electron microscopy. Two-way ANOVA revealed that conditioning treatment and interaction between treatment and dentin depth significantly influenced shear bond strength results. Acid-etching alone yielded shear bond strength values that were significantly higher than those achieved with laser ablation alone, or in combination with acid-etching. The Er-YAG laser created a laser-modified layer that adversely affects adhesion to dentin, so it does not constitute an alternative bonding strategy to conventional acid etching.