Aim: To express and purify human MT-2a in prokaryotic cells and to prepare the MT-2a-specific rabbit antiserum.
Methods: GST-MT-2a fusion protein was expressed after IPTG induction and further purified with Glutathione Sepharose 4B. Then the purified GST-MT-2a fusion protein was used to immunize New Zealand rabbits. The titer and specificity of rabbit antiserum were evaluated by double immunodiffusion, ELISA and Western blot.
Results: GST-MT-2a fusion protein was highly expressed. The final yield of the pure GST-MT-2a was about 38 mg per liter of bacterial culture. Its antiserum with high specificity and potency was also obtained.
Conclusion: The successful expression of GST-MT-2a fusion protein in E. coli and the preparation of MT-2a specific rabbit antiserum will be valuable for the study on the function of human MT-2a.