In this research, a high performance, ionomer-free electrocatalyst based on vertically aligned palladium (Pd) nanowire array was developed as an anode electrode toward ethanol oxidation reaction (EOR) in an alkaline environment. Using a one-step electrodeposition method, the Pd nanowires with controlled length were obtained by varying the electrodeposition current density and the synthesis time. Scanning electron microcopy (SEM), energy dispersive X-ray spectroscopy (EDS), and X-ray powder diffraction (XRD) were employed to characterize the morphology, chemical composition, and crystal structure of the Pd nanowires. The length effects of the nanowires, in the range of 0.8–4.5 μm, and various metal substrates, such as Ag, Cu, Ni, and Ti, were investigated for their electrochemical activities. The results demonstrated that Ag was the most active substrate to facilitate the ethanol oxidation reaction of the Pd nanowire array (NWA) electrocatalyst, which could be related to its good electrical conductivity. The stability test of the Pd NWA/Ag over time for EOR was also carried out, and the catalytic activity was recovered after the electrode was replaced with a new ethanol solution. Electrochemical impedance spectroscopy (EIS) measurements were performed to provide insights in the electron transfer resistance between the electrode and analyte. Gas chromatography and UV–vis spectroscopy were employed to measure the concentration of chemical species, which helped elucidate the overall reaction mechanism on the electrode surfaces.