A newly developed passive device that provides negative stiffness has been implemented within a quarter-scale highway bridge model and subjected to seismic loading via shake table testing. Details of the experimental results and their comparison with numerical simulations under a wide range of ground motions are presented. In addition, performance indices were developed to systematically evaluate the relative performance of different isolation system configurations that employ combinations of positive and negative stiffness as well as various levels of damping. Further, the influence of boundary conditions (rigid versus flexible bridge piers) on the effectiveness of employing negative stiffness devices has been evaluated.