Preparation and Purification of Anti-human DR5 Polyclonal Antibody

Objective: To purify GST-eDR5 fusion protein and prepare the mouse anti-human DR5 antisera.Method: The recombinant protein GST-eDR5 was purified by GST purification kit and PAGE.The polyclonal antibody against GST-eDR5 fusion protein was prepared by immunizing mouse with the purified GST-eDR5 fusion protein,and then was purified by reaction with GST.The specificity and titer of polyclonal antibody were detected by western blotting and indirect ELISA.Result: High purified fusion protein GST-eDR5 was obtained and its concentration is 0.65μg/μl.Very high specificity of mouse anti-human DR5 antiserum was generated and analyzed by western blotting.The titer was as high as 1∶25600.The antisera purified by GST fails to recognize GST and only recognize native DR5 from cells and the recombinant GST-eDR5 fusion protein.Conclusion: The polyclonal anti-DR5 antibody with high specificity and high titer has been generated successfully.This study settles a foundation for investigating its inhibition and apoptosis effect on tumor cells.