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高等学校化学研究 ›› 2006, Vol. 22 ›› Issue (3): 308-311.

• Articles • 上一篇    下一篇

Preparation of Polyclonal Antibodies Against Testis-specific Protease 50 and Characterization of Antibody Specificity

ZHANG Jing1, YANG Ming1, XU Ze-Li2, BAO Yong-li1, WU Yin1, WANG Yue-zeng1, MENG Xiang-ying3, JU Xing-da1, GU Yang1, LI Yu-xin3   

  1. 1. Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, P. R. China;
    2. China-Japan Union Hospital, Jilin University, Changchun 130031, P. R. China;
    3. Research Center of Agriculture and Medicine Gene Engineering of Ministry of Education
  • 收稿日期:2005-11-25 出版日期:2006-06-24 发布日期:2011-08-06
  • 基金资助:

    Supported by the Distinguished Young Scholars Fund of Jilin Province(No. 20050114).

Preparation of Polyclonal Antibodies Against Testis-specific Protease 50 and Characterization of Antibody Specificity

ZHANG Jing1, YANG Ming1, XU Ze-Li2, BAO Yong-li1, WU Yin1, WANG Yue-zeng1, MENG Xiang-ying3, JU Xing-da1, GU Yang1, LI Yu-xin3   

  1. 1. Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, P. R. China;
    2. China-Japan Union Hospital, Jilin University, Changchun 130031, P. R. China;
    3. Research Center of Agriculture and Medicine Gene Engineering of Ministry of Education
  • Received:2005-11-25 Online:2006-06-24 Published:2011-08-06
  • Supported by:

    Supported by the Distinguished Young Scholars Fund of Jilin Province(No. 20050114).

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摘要： Testis-specific protease 50 (TSP50) is a testis-specific oncogene, which is abnormally activated in most tested patients with breast cancer. This property makes it an attractive molecular marker and a promising target for the diagnosis and therapy of breast cancer. In order to obtain the protective and specific polyclonal antibodies for further research, TSP50 cDNA was amplified by RT-PCR from normal human testicular tissue, and inserted into eukaryotic expression vector pcDNA3.1. Rabbit anti-TSP50 polyclonal antibodies were prepared by means of intramuscular injection of pcDNA3.1-TSP50 into the rabbits. Titers of the anti-sera were measured by ELISA and Western blotting with the E. coli cell lysate containing the induced GST-TSP50 fusion protein as an antigen. In addition, we examined the expression of TSP50 in both breast cancer cell line MCF-7 and breast cancer tissue by immunofluorescent and immunohistochemistry analysis.

关键词: Breast cancer, ELISA, Polyclonal antibody, TSP50

Abstract: Testis-specific protease 50 (TSP50) is a testis-specific oncogene, which is abnormally activated in most tested patients with breast cancer. This property makes it an attractive molecular marker and a promising target for the diagnosis and therapy of breast cancer. In order to obtain the protective and specific polyclonal antibodies for further research, TSP50 cDNA was amplified by RT-PCR from normal human testicular tissue, and inserted into eukaryotic expression vector pcDNA3.1. Rabbit anti-TSP50 polyclonal antibodies were prepared by means of intramuscular injection of pcDNA3.1-TSP50 into the rabbits. Titers of the anti-sera were measured by ELISA and Western blotting with the E. coli cell lysate containing the induced GST-TSP50 fusion protein as an antigen. In addition, we examined the expression of TSP50 in both breast cancer cell line MCF-7 and breast cancer tissue by immunofluorescent and immunohistochemistry analysis.

Key words: Breast cancer, ELISA, Polyclonal antibody, TSP50