Galectin-1, a mammalian galactoside lectin is present in many cells and tissues. It is involved in many physiological processes, such as cell adhesion, proliferation, apoptosis, and inflammation. Galectin-1 is a member of the β-galactoside binding family, which are involved in the regulation of the immune system and development of tumors. Antler is a rare accessory organ that can periodically shed and regenerate from its pedicle in deer and has attracted attention as a new model for studying mammalian organ regeneration. Deer antlers offer a unique opportunity to explore how nature has devised a system of epimorphic regeneration in mammals. Antler regeneration is a stem cell-based process derived from antler stem cells that are located in the pedicle periosteum (PP). Elevated expression of galectin-1 in antler stem cells suggest that this molecule may play an important role in antler regeneration. To study the biological function of Galectin-1 in antler regeneration, it is necessary to generate a polyclonal antibody. In this study, the Galectin-1 gene of Sika deer (Cervus nippon) was inserted into the pET28a vector to obtain the recombinant plasmid, pET28a-Galectin-1. This recombinant plasmid was then transformed into Escherichia coli strain BL21(DE3) for induction and expression. The target protein was purified by using a Ni-NTA agarose affinity column. The purified protein was used to immunize rabbits to obtain the polyclonal antibody. Antibody titer was determined by enzyme-linked immunosorbent assay (ELISA) for Galectin-1. The specificity of antibody was confirmed by western blot analysis and the expression of Galectin-1 in the antler stem cells was detected by an immunofluorescence assay. The results showed that the titer of the antibody was 1:64000 with high specificity and that Galectin-1 was expressed in almost all of the antler PP cells. We have generated a specific polyclonal antibody of deer Galectin-1, which will serve as an important tool for the study of the regulation of Galectin-1 in antler regeneration.