Canine parvovirus (CPV), canine distemper virus (CDV) and rabies are viruses that cause life-threatening infections in dogs and have a large effect on the canine population. This work aims to investigate the effect of inactivated rabies vaccine on the immune response of vaccinated puppies with live attenuated CD and CP vaccines. Antibody response of puppies vaccinated with CD, CP and rabies vaccines either in single or simultaneous administration was evaluated through vaccination of different puppy groups and monitoring the levels of induced antibodies in their sera through application of SNT and indirect ELISA. The two serological assays revealed that the simultaneous vaccination of puppies with CD or CP vaccines with rabies vaccine induced higher levels of CD or CP antibodies 256 by SNT and 3.5 log10 by ELISA) than in case of Puppies vaccinated with live CD or live CP vaccines alone. So, it could be recommended to vaccinate puppies simultaneously with attenuated CD and inactivated rabies vaccines, or attenuated CP and inactivated rabies vaccines or with the three vaccines at one shot.