Abstract: To standardize the indirect ELISA under local conditions, antispecies-antibodies were produced in rabbits. The antibodies against Pasteurella multocida were produced in buffalo calves by injecting the formalin, heat-killed organism_ These antibodies were inoculated into rabbits to produce rabbit-antibuffalo antibodies. These were isolated and partially purified by ammonium sulfate precipitation technique. Protein contents were estimated using Biuret method and the presence of rabbit-antibuffalo antibodies was confirmed and measured through agar gel precipitation test (AGPT). These isolated anti-antibodies were conjugated with horseradish peroxidase by two-step glutaraidehyde method. Ten mg of horseradish peroxidase was applied for the Anti-antibody-Enzyme Conjugate formation and the best results were recorded at 1 :100 and 1.200 dilutions of above conjugate when tested through indirect ELISA for H.S. anti-antibodies.