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学科主题: 基础医学
题名:
Expression, purification and identification of a novel human cytokine C-terminal 51 amino protein of CKLFSF2 in E-coli
作者: Zhong Ying-Cheng; Shi Shuang; Zhang Ying-Mei; Mo Xiao-Ning; Liu Da-Zhen; Song Quan-Sheng; Han Wen-Ling; Wang Ying
关键词: CKLFSF2 (chemokine-like super family member 2) ; recombinant protein purification ; polycolonal antibody ; chemotaxis ; immunohistochemistry ; antibody neutralization
刊名: PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS
发表日期: 2007
卷: 34, 期:1, 页:93-99
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemistry & Molecular Biology ; Biophysics
研究领域[WOS]: Biochemistry & Molecular Biology ; Biophysics
关键词[WOS]: CHEMOKINE-LIKE FACTOR-1 ; MOLECULAR-CLONING
英文摘要:

Overexpressed CKLFSF2 (chemokine-like super family member 2) can be secreted into the supernatant of cultured cells, which exhibits the effects on cell-chemotaxis. To study the structural and functional characteristics of CKLFSF2 and make the polycolonal antibody, the recombinant plasmid pGEX-4T-3-CKLFSF C51 was constructed, following the expression in E. coli with high efficiency, purification using Glutathione Sepharose 4B and Sephacyl S-200, the recombinant CKLFSF2 C51 protein (typically > 95% pure) was obtained. The N-terminal animo acid sequencing of CKLFSF2 C51 was performed and relative molecular mass is 5871.48 measured by ASIMA-CFR (TM) plus MALDI-TOF. ELISA assay detected the reactivity of the polycolonal antibody and Western blot show that CKLFSF2 was overexpressed in mammalian cells and the specific band was corresponding to the expectation. The recombinant CKLFSF2 C51 protein exhibits significantly chemotactic effect in PC-3 cells, which can be neutralised by anti-CKLFSF2 C51 polyantibody. The anti-CKLFSF2 C51 antibody can also be used for immunohistochemistry. Taken together, the recombinant CKLFSF2 C51 from E.coli has similar bioactivity as that from eukaryocytic expression system. The polycolonal antibodies of recombinant CKLFSF2 C51 can be used in immunohistochemistry, Western blot and to neutralize the chemotactic effect.

语种: 中文
WOS记录号: WOS:000243624800013
Citation statistics:
内容类型: 期刊论文
版本: 出版稿
URI标识: http://ir.bjmu.edu.cn/handle/400002259/60784
Appears in Collections:基础医学院_免疫学系_期刊论文

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人类新细胞因子CKLFSF2羧基_省略_杆菌中的表达纯化_抗体制备与鉴定_钟英成.pdf(316KB)期刊论文出版稿限制开放View Download

作者单位: Peking Univ, Hlth Sci Ctr, Lab Med Immunol, Sch Basic Med Sci, Beijing 100083, Peoples R China

Recommended Citation:
Zhong Ying-Cheng,Shi Shuang,Zhang Ying-Mei,et al. Expression, purification and identification of a novel human cytokine C-terminal 51 amino protein of CKLFSF2 in E-coli[J]. PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS,2007,34(1):93-99.
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文件名: 人类新细胞因子CKLFSF2羧基_省略_杆菌中的表达纯化_抗体制备与鉴定_钟英成.pdf
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