The purpose of this in vitro study was to evaluate the effect of immediate dentin sealing (IDS) with and without chlorhexidine (CHX) pretreatment on the shear bond strength (SBS) of dual-cure adhesive resin cement. Mid-coronal dentin surfaces were obtained from 75 human molars. They were randomly allocated into five groups in accordance to type of IDS [etch&rinse/ER (Adper Single Bond2) and universal/U adhesive systems (Single Bond Universal)] and presence of CHX application (n = 15): Group ER; Group ER + CHX; Group U; Group U + CHX; and Group C (no IDS). Dual-cure adhesive resin cements were bonded with a cylinder-shaped Teflon mold. The SBS was measured using a universal test machine. Fracture type was evaluated with stereomicroscope. The resin/dentin interfaces were examined with an environmental scanning electron microscope. Data were statistically analyzed with two-way analysis of variance and Bonferroni tests (p < .05). Regarding the IDS treatment groups with/without CHX, there were no significant differences in SBS (p > .05). Group U + CHX showed significantly higher SBS than Group C (p < .05). Regarding the presence of CHX, no significant differences in SBS were found (p > .05). The prevalent failure mode was the mixed type for most of the groups. Group C exhibited an intact and regular hybrid layer with no resin tag, whereas longer and clear resin-tag formation was visible for Group U + CHX. CHX pretreatment improved the bond strength between adhesive resin cement and dentin when IDS treatment was performed with a universal adhesive system.