Preparation and Purification of Polyclonal Antibody Against N-Terminal of ATP7B

To prepare and purify specific antibody against N Terminal of ATP7B(Wilson Protein),and to make a good foundation for study of the structure and function of ATP7B. The target gene was amplified by RT PCR. Fusion protein was expressed with GST gene fusion system. Affinity chromatography was used for protein purification. Thrombin was used to cleave fusion protein with on column cleavage method. The aimed protein was gathered as antigen. Rabbits were immunized subcutaneously. The titre of the antibody was detected by ELISA assay. The antiserum was purified with DEAE Sephadex A 25 columns. Western blot was applied to confirm the specificity of the antibody.ELISA assay revealed that the titer of the prepared antiserum against fusion protein was as high as 1∶2 500. Western blot showed that the antibody was able to react with the proteins. Non denatured SDS PAGE confirmed high purity of IgG. [Conclusion]Polyclonal antibody against ATP7B,is successfully prepared.