[Preparation and application of polyclonal antibody of human transcription factor CTCF N terminus protein].
To investigate the function of CTCF and understand the pathogenesis of tumors better, we produced rabbit polyclonal antibody of human transcription factor CTCF protein and detected its expression in several kinds of human cancer cells and tissues. GST fusion protein of human CTCF-N domain was purified by GSTrap-FF affinity chromatography and was successfully expressed under induction of IPTG in E. coli BL21. Western blotting analysis demonstrated that the polyclonal antibody can recognize the endogenous CTCF from HepG2, MCF-7 and HeLa cells specifically. The produced antibodies can be used for gene expression regulation and tissue distribution study at protein level.
Zhang
Yingchun
Y
Department of Biochemistry and Molecular Biology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Jiang
Lei
L
Wei
Ling
L
Chai
Xinjuan
X
Ge
Yajun
Y
Qin
Yang
Y
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000378:metabolism*; D000076246:CCCTC-Binding Factor; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D012097:Repressor Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*
D000906:Antibodies; D000076246:CCCTC-Binding Factor; C000619697:CTCF protein, human; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D005982:Glutathione Transferase