The electrodeposition of reduced graphene oxide (ERGO) film with preferred vertical orientation was fabricated on a glassy carbon electrode by using pulse potential method in a graphene oxide colloidal solution. Using square wave adsorptive stripping voltammetry (SWASV), the ERGO film was applied for the first time, in developing a high-sensitive electrochemical sensor for detection of taxifolin. Compared with bare glassy carbon electrode (GCE), the resulting electrodes (ERGO/GCE) exhibited excellent response toward the redox of taxifolin by significantly enhancing the redox peak currents and decreasing the peak-to-peak separation. Under the selected conditions, the peak currents were linear relationship with taxifolin concentration in the range of 1.0 × 10−8–1.0 × 10−6 mol L−1, with detection limit of 2.0 × 10−9 mol L−1. Besides, the ERGO/GCE also exhibited an excellent selectivity, stability, reproducibility and repeatability.