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学科主题: 临床医学
题名:
Preparation and characterization of monoclonal antibodies against nucleostemin, a protein that controls cell proliferation in stem cells and cancer cells
作者: Yang, HX; Zhang, JZ; Wu, J; Shou, CC
刊名: HYBRIDOMA
发表日期: 2005-02-01
卷: 24, 期:1, 页:36-41
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemical Research Methods ; Biotechnology & Applied Microbiology ; Immunology
研究领域[WOS]: Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Immunology
英文摘要:

Nucleostemin (NS) is a novel protein that promotes the proliferation of stem cells and cancer cells. The aim of this study is to prepare specific monoclonal antibodies against nucleostemin as a tool in nucleostemin research. To generate the antibodies, the cDNA coding nucleostemin was cloned from human gastric cancer cell line NCI-N87 with reverse transcriptase-polymerase chain reaction (RT-PCR). The partial cDNA fragment was inserted into prokaryotic expression vector pGEX-4T1 to make glutathione S-transferase (GST) fusion protein in Escherichia coli, and the whole-length cDNA fragment was cloned into vector pcDNA3 for expression in mammalian cells. The spleen cells, from the BALB/c mouse immunized with the GST-nucleostemin fusion protein, were used to prepare the monoclonal antibody with hybridoma technique. After screening with enzyme-linked immunosorbent assay (ELISA) and subcloning, six specific monoclonal antibodies against nucleostemin were obtained. Moreover, they also worked well in Western blotting, immunoprecipitation, and immunohistochemistry. These antibodies will be helpful in elucidating biological functions and the molecular mechanism of nucleostemin.

语种: 英语
WOS记录号: WOS:000228474200005
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/50381
Appears in Collections:北京大学临床肿瘤学院_期刊论文

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作者单位: Peking Univ, Sch Oncol, Beijing Inst Canc Res, Dept Biochem & Mol Biol, Beijing 100034, Peoples R China

Recommended Citation:
Yang, HX,Zhang, JZ,Wu, J,et al. Preparation and characterization of monoclonal antibodies against nucleostemin, a protein that controls cell proliferation in stem cells and cancer cells[J]. HYBRIDOMA,2005,24(1):36-41.
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