Background and Aim : In the recent century use of radiofrequency electromagnetic radiation emitted from Wi-Fi equipment has increased. Using this device in houses, workplaces, public areas, schools have become a routine task in our daily lives, which consequently can affect various systems of human body. Change in the levels of energy and spin orientation of electrons under electromagnetic fields can increase the production of reactive oxygen species (ROS). The increase of reactive oxygen species may affect the enzymatic antioxidant systems such as Glutathion S-transferase in blood. The present study was designed to determine the possible effect of electromagnetic radiation emitted from Wi-Fi equipment on Glutathion S-transferase level in rat blood. Methods : In this study twelve male rats were selected and divided into two groups of case and control, six in each group. On the 10th week of the study, blood samples were taken and used to assay Glutathion S-transferase level. Results : : Level of enzyme in rat plasma significantly increased in Wi-Fi exposed rats (80.19±18.09 Vs 50.87±20.62, p=0.026). Conclusion : In conclusion, changes in Glutathion S-transferase level may be due to increased level of free radicals after exposure to radiofrequency radiation emitted from Wi-Fi equipment. Therefore, reinforcement of antioxidant defense system may be required to eliminate free radicals produced under this condition.