We demonstrate the effect of the depletion force in experiments and simulations of vertically vibrated mixtures of large and small steel spheres. The system exhibits size segregation and a large increase in the pair correlation function of the large spheres for short distances that can be accurately described using a combination of the depletion potential derived for equilibrium colloidal systems and a Boltzmann factor. The Boltzmann factor defines an effective temperature for the system, which we compare to other measures of temperature.