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Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:

2014 01

Page:

29-0033

Research Field:

Publishing date:

Info

Title:

Preparation and application of anti-mouse fibroblast activation protein polyclonal antibody

Author(s):

Jiang Guanmin¹; Xie Wanying¹; Zhang Kunshui²; Qiu Yu³; Zhang Qiugui¹; Du Jun⁴

1.Department of Clinical Laboratory,The First Affiliated Hospital of University of South China,Hunan Hengyang 421001,China;2.Out-patient Department of South Campus,The Sun Yat-sen Memorial Hospital of Sun Yat-sen University,Guangdong Guangzhou 510120,China;3.Department of ICU,Hunan Children's Hospital,Hunan Changsha 410007,China;4.Laboratory of Microbial and Biochemical Pharmacy,School of Pharmaceutical Sciences,Sun Yat-sen University,Guangdong Guangzhou 510006,China.

Keywords:

fibroblast activation protein; polyclonal antibody; tumor metastasis; tumor stroma

PACS:

R730.3

DOI:

10.3969/j.issn.1672-4992.2014.01.09

Abstract:

Objective:To express and purify His-FAP¦Ác fusion protein and to generate rabbit anti-mouse FAP¦Á polyclonal antibody,which was used to analyze FAP¦Á expression in tumor cells,Fibroblast cells and tumor stroma.Methods:A prokaryotic expression plasmid of pET28a(+)-FAP¦Ác constructed by our laboratory was transformed into BL21.The expression of His-FAP¦Ác protein was induced by IPTG.The anti-mouse FAP¦Á polyclonal antibody was obtained by immunizing rabbits with purified His-FAP¦Ác protein.The quality of the antibody was identified by Western blot and ELISA.Whether FAP¦Á expression in mouse tumor cells,fibroblast cells and tumor stroma was also analyzed using the antibody.Results:His-FAP¦Ác fusion protein was specifically combined with rabbit anti-mouse FAP¦Á polyclonal.The polyclonal antibody was of high titer with high specificity.It could recognize FAP¦Á expression in mouse embryo fibroblast cells and tumor stroma.Conclusion:The rabbit anti-mouse FAP¦Á polyclonal antibody could recognize FAP¦Á expression in mouse embryo fibroblast cells and tumor stroma.Therefore,providing a tool to study the role of FAP¦Á in tumor generation,promotion and metastasis.It also establishes a foundation of further anti-tumor research.

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