Abstract: Objective: To prepare the anti-LRP16 polyclonal antibody and investigate the subcellular localization of LRP16 protein. Methods: The full length LRP16 gene was obtained by RT-PCR. The predicated antigen epitope was cloned into protein expression plasmid pRSET-C, transformed into E.coli BL21（DE3） cells for expressing recombinant pRSET-C-LRP16 protein induced by IPTG. The purified pRSET-C-LRP16 protein was employed to immunize rabbit for preparing the polyclonal antibody. The expression of LRP16 was analyzed by Western blot and immunohistochemistry in cancer such as: breast cancer and ovarian serous cystadenoma. Results: Specific anti-LRP16 rabbit polyclonal antibody was obtained, and both Western blot and immunohistochemistry showed that LRP16 was expressed in the cancer with high-expressing of ER, localized in the cellular nucleus. Conclusions: LRP16 protein is a nuclear protein located in cellular nucleus. The anti LRP16 polyclonal antibody can be used for studying the physiological function of LRP16 and its pathology.