In order to study the effects of adhesive, substrate material and ambient temperature on the mechanical properties of the adhesive joint, the quasi-static tensile mechanical properties of the single lap adhesive joint were studied. The results show that the bonding strength of epoxy resin adhesive is better than that of polyurethane adhesive, but the polyurethane adhesive shows good ductility. The differences of substrate materials would affect the strength of the adhesive joint. The plastic deformation at the lap end of aluminum substrate would accelerate of the failure process of adhesive layer, leading to the reduction of the joint strength. With the increase of temperature, the strength and stiffness of the adhesive joint decreases, but the decrease is related to the type of adhesive. It is mainly related to the glass transition temperature of the adhesive. In addition, the failure mode of the joint is affected by the ambient temperature. At high temperature, the joint is more prone to adhesive failure. The study provides a more comprehensive guidance for the application of single lap adhesive joint in the actual service environment of automobiles.