In this study, the immunogenicity of a rabies vaccine that was used in one of the Isfahan cattle farms is reported. This vaccine was produced by replication of the Pasteur virus (PV) strain on BHK-21 monolayer cell culture. The virus suspension was inactivated by ß-propiolactone treatment and the harvested pool showed sterility and safety. Thus, the obtained vaccine was tested on mice and a satisfactory immune response was observed. Thereafter, the vaccine was tested on 34 cattle divided into 2 groups. Each group was received a different dose of the vaccine intramuscularly. Sera were taken 2 months after rabies vaccination and were analyzed by the rapid fluorescence focus inhibition test (RFFIT) to evaluate the titer of the rabies-neutralizing antibody. Both groups under study showed a sufficient seroconversion.