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人CTCF N端融合蛋白的表达、抗体制备及鉴定 被引量：1

Preparation and Application of Polyclonal Antibody of Human Transcription Factor CTCF N Terminus Protein
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摘要 为研究CTCF的功能,进一步探索肿瘤的发病机制,我们成功制备了人转录因子CTCF N端融合蛋白多克隆抗体,并初步用于肿瘤细胞CTCF表达状态的分析。采用已构建的表达质粒PGEX-4T-2-CTCF-N,经IPTG诱导表达GST-CTCF N融合蛋白,亲和层析纯化GST-CTCF N融合蛋白,免疫新西兰大白兔制备多克隆抗体。并通过Western blot,对肿瘤细胞和组织CTCF蛋白表达状态进行初步分析。本实验成功在大肠杆菌BL21中诱导表达了GST-CTCF N融合蛋白,经亲和层析纯化获得了纯化的融合蛋白。免疫新西兰大白兔获得了较高生物活性和特异性的多克隆抗体,Western blot证实此多克隆抗体能够特异性识别HepG2、HeLa、MCF-7癌细胞及组织的内源性CTCF蛋白,为今后从蛋白质水平研究CTCF基因表达调控研究打下了基础。 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.
作者 张迎春 蒋磊 魏玲 柴新娟 葛亚俊 覃扬
机构地区 四川大学华西医学中心生物化学与分子生物学教研室
出处 《生物医学工程学杂志》 EI CAS CSCD 北大核心 2010年第2期379-383,共5页 Journal of Biomedical Engineering
基金 国家自然科学基金资助项目(30870957)
关键词 CTCF 多克隆抗体 WESTERN BLOT 肿瘤细胞表达 CTCF Polyclonal antibody Western blot Cancer cellular expression
分类号 R392 [医药卫生—免疫学]
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参考文献12

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二级参考文献24

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共引文献5

同被引文献19

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生物医学工程学杂志
2010年 第2期

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