Graphene oxide membranes were prepared by vacuum and pressurized ultrafiltration methods on the 12% modified Polyacrylonitrile (12mPAN) substrate to specify challenges, salient features, future directions, and potential of GO membrane for separation fields using characterization techniques and gas separation test (studied gases are CO2, He and N2), which is an efficient tool for better understanding of GO membrane behavior. GO membrane structure was examined over a wide range of parameters, such as pore size range of substrate and its surface properties, pH of GO dispersion, GO content, synthesis pressure, operating pressure and temperature. The results show that the GO content does not hold a linear relationship with the permeance and selectivity. Film thickness, aggregates, synthesis pressure defects and interlayer spacing have significant effects on the gas separation performance of GO membranes which originate from the synthesis method and its conditions.