Objective To express and purify TDG protein and prepare the rabbit antibody against thymine DNA glycosylase (TDG) protein. Methods　Mouse TDG (mTDG) was amplified by PCR, and then was inserted into the fusion expression vector pET28a(+). After being expressed in E. coli BL21, the His-mTDG protein was purified and used to immunize rabbit. Purified antibody was obtained through affinity chromatography column，and its Applications was evaluated through western blot and immunofluorescence analsysis. Results　 His-mTDG fusion protein was almost all soluble at low temperature (20°C) and low IPTG condition (0.2 mM). After the fusion protein was purified and immunized rabbits, the antibody against TDG was obtained. The further results showed that the antibody had a good specificity in immunoassays. Conclusion　we have purified mTDG fusion protein with an obviously homogeneous band and high biology activity and prepared the rabbit antibody against TDG successfully, which lays the foundation for further study of it’s character, structure and function.