After the Fukushima Daiichi Nuclear Power Plant disaster in Japan in 2011, the demand drastically increased for efficient technology for the removal of radioactive cesium. Prussian blue (PB) nanoparticles have shown excellent adsorption ability toward Cs. In this study, we synthesized PB nanoparticles incorporated polyacrylonitrile nanofiber (PB/PAN). PB/PAN has the porous structure of nanofibers, with diameters of several hundred nanometers. PB nanoparticles can be incorporated successfully into the PAN matrix without any change to their intrinsic crystallinity and structure. The mesoporous structure of PB/PAN and the incorporation of PB nanoparticles led to an increase in the Brunauer-Emmett-Teller surface area and pore volume. In addition, PB/PAN exhibited excellent wettability with water. With simple filtering for the removal of radioactive cesium, PB/PAN showed high removal efficiency (87 ± 3%) within 10 s for 10 mL of Cs solution (1000 Bq L). In addition, the Cs removal by PB/PAN showed high removal efficiency (70 ± 2%, after 1 h), even in the actual seawater medium (1000 Bq L of Cs). Therefore, PB-incorporated PAN nanofibers can be considered useful in the practical application of Cs removal from radioactive wastewater.