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学科主题基础医学
Expression, purification and identification of a novel human cytokine C-terminal 51 amino protein of CKLFSF2 in E-coli
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
2007
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
引用统计
文献类型期刊论文
版本出版稿
条目标识符http://ir.bjmu.edu.cn/handle/400002259/60784
专题基础医学院_免疫学系
作者单位Peking Univ, Hlth Sci Ctr, Lab Med Immunol, Sch Basic Med Sci, Beijing 100083, Peoples R China
推荐引用方式
GB/T 7714
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.
APA Zhong Ying-Cheng.,Shi Shuang.,Zhang Ying-Mei.,Mo Xiao-Ning.,Liu Da-Zhen.,...&Wang Ying.(2007).Expression, purification and identification of a novel human cytokine C-terminal 51 amino protein of CKLFSF2 in E-coli.PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS,34(1),93-99.
MLA Zhong Ying-Cheng,et al."Expression, purification and identification of a novel human cytokine C-terminal 51 amino protein of CKLFSF2 in E-coli".PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS 34.1(2007):93-99.
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