Preparation of polyclonal antibody against Stat3 and its application in the diagnosis of breast cancer

    LI Hongª²Fen, SHAN Baoª²En, CHEN Jing, DONG Qing, WANG Xiaoª²Ling, WANG Yongª²Jun, CHEN Xing, DONG Jinª²Zhuo, MA Hong

    Research Center, Fourth Hospital, Hebei Medical University, Bethune International Peace Hospital, Shijiazhuang 050011, China, American MD Corporation

    ¡¾Abstract¡¿ AIM: To prepare rabbit antibody against Stat3 and analyze its application in the diagnosis of breast cancer. METHODS: The male rabbits were immunized with recombinant mouse Stat3 protein to prepare antibody against Stat3. The purity and specificity of the antibody were analyzed by Western Blot. The expression of Stat3 protein in the 65 cases of breast cancer and 30 cases of normal breast tissue was detected by immunohistochemical staining. RESULTS: The titer of antiª²Stat3 antibody in serum was >1¡Ã256 000 (A450=0.40) at the 6th week after the immunization. Both SDSª²PAGE and Western Blot analysis showed that there was an obvious protein band with Mr 92 000, The intracytoplasmic expression levels of Stat3 protein in the different histological gradings of 65 cases of breast cancer showed significant difference (P<0.05), so did the intranuclear expression levels (P<0.05) . CONCLUSION: Polyclonal antibody against Stat3 with high titer and specificity can be prepared successfully, and it can be used to detect the expression of Stat3 protein in breast cancer tissue. ......