An Ag2O/Ag electrode was prepared through the electrochemical oxidation of sterling silver. This electrode was used as a cathodic electron acceptor in a microbial fuel cell (MFC). The Ag2O/Ag electrode was characterized by scanning electron microscopy, X-ray powder diffraction and linear sweep voltammetry. The maximum voltage output of the MFC with the Ag2O/Ag cathode was maintained at between 0.47 and 0.5 V in 100 cycles, indicating the good regenerative capacity of the Ag2O/Ag electrode. The overpotential loss for silver oxide was 0.021–0.006 V, and the maximum power output, open circuit potential and short circuit current of the MFC were 1.796 W m−3, 0.559 V and 9.3375 A m−3, respectively. The energy required for electrochemical reoxidation ranged from 40% to 55% of the energy produced by the MFC. Results indicated that the Ag2O/Ag electrode could be used as a cathodic electron acceptor in MFCs with excellent stability.