The poly(o-anisidine)-sulfuric acid-glucose oxidase (POA-H2SO4-GOx) electrode has been investigated in the present work. Platinum electrode was used for the synthesis of poly (o-anisidine)-sulfuric acid (POA-H2SO4) film using galvanostatic method with 0.2 M o-anisidine, 1.0 M H2SO4 solution, 1.0 pH and 2 mA/cm(2) applied current density. The synthesized film was characterized using electrochemical technique, conductivity measurement, UV-visible spectroscopy, Fourier transform infrared spectroscopy, and scanning electron microscopy. GO(X) was immobilized on synthesized POA-H2SO4 film by cross-linking via glutaraldehyde in phosphate and acetate buffer. The Michaelis-Menten constant (K'(m)) was determined for the immobilized enzyme. The glucose oxidase electrode shows the maximum current response at pH 5.5 and potential 0.6 V. The sensitivity of POA-H2SO4-GO(X) electrode in phosphate and acetate buffer has been recorded. The results of this study reveal that the phosphate buffer gives fast response as compared to acetate buffer in amperometric measurements.