Objective To prepare the rabbit polyclonal antibodies against seventeen mammals and to label the secondary antibodies with horseradish peroxidase for developing the serological test system of mammals． Methods IgGs of mammals were purified by precipitation with ammonium sulfate combining affinity chromatography using Protein A or Protein G． Rabbit anti-sera were prepared by repeatedly immunized rabbits with purified IgGs． The antibodies against mammal IgG in rabbit sera were detected by agarose double immunodiffusion and purified using Protein A affinity chromatography． The products were labeled using HRP by periodate oxidation method． And the activities of the labeled rabbit polyclonal antibodies against seventeen mammals were detected by ELISA． Western blots were performed to investigate the specificity of the seventeen rabbit anti-mammal IgGs-HRP． Results The IgGs from the seventeen mammal species were purified，including rhesus，tiger，brandt's vole，striped hamster，goral，guanaco，civet cat，crab-eating macaque，sika deer，mongolian gerbil，red deer，camel，hog deer，ibex，hamster，assam macaque，ferret． The titers of seventeen rabbit antisera were up to 1 ∶ 32 examined by agarose double immunodiffusion． Seventeen kinds of purified rabbit anti-mammalsecondary antibodies were labeled with HRP and their activities were up to 1 ∶ 2000 － 60000 examined by ELISA． Thespecific activities of the seventeen rabbit anti-mammal IgGs-HRP were indicated with western blot． Conclusion Rabbit polyclonal antibodies against seventeen mammals labeled with HRP were prepared successfully，it is an useful tool for developing the serological test system of mammals．