MAP法制备抗人Connexin31抗体

The Antiserum of the Multiple Antigenic Peptide of the Connexin31:Preparation and Application

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摘要 MAP(multipleantigenicpeptide)多肽法具有快速、简便、成本低廉的特点.用MAP法,直接在多肽合成仪上合成Connexin31羧基端一个多肽片段(250-266AA)的MAP肽,经超滤器纯化后,免疫新西兰雄兔后采血检测、并纯化.经Westernblotting、细胞免疫荧光染色、免疫沉淀证实得到的抗体为抗间隙连接蛋白31的特异抗体. MAP peptide has been shown to produce strong immunogenic response. Synthesized the multiple antigenic peptide, then the purified multiple antigenic peptide solution was mixed with Freund＇ s complete or incomplete adjuvant and immunized rabbit to prepare specific polyclonal antibody against the topology of Connexin31. A full-length human Connexin31 gene was cloned into expression vector, pcDNA3.1/myc-his （-） B, and transformed into HTI080 cell line stably or into Hela cell transiently. Western blotting analysis and immunoprecipitation suggested that the polyclonal antibody recognize the fusion protein of the Connexin31-myc. Cell immunofluorescence staining shows the Connexin 31 localized at the site of the cell-cell contact gap junction. The antiserum against Connexin31 prepared by multiple antigenic peptide has good specificity.

作者刘宇谭志平蒋泰文贺力强胡艺俏姚仲元蔡芳夏昆夏家辉

机构地区湖南理工学院化学化工系中南大学医学遗传学国家重点实验室

出处《生命科学研究》 CAS CSCD 2005年第4期308-312,共5页 Life Science Research

基金国家863项目(2002BA711A07) 国家自然科学基金资助项目(30370737 39970372)

关键词间隙连接蛋白31 MAP肽免疫荧光免疫印迹免疫沉淀 Connexin3 l multiple antigenic peptide immunofluorescence Western blotting immunoprecipitation

分类号 Q25 [生物学—细胞生物学]

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1刘宇,蔡芳,谭志平,贺立强,蒋泰文,黄亮群,夏昆,夏家辉.一种特异性多用途抗人Connexin31抗体的获得与应用[J].激光生物学报,2005,14(2):81-86.
2刘宇,寻晓红,聂东宋.抗人Connexin31单克隆抗体的获得与应用[J].湖南理工学院学报（自然科学版）,2006,19(3):71-73.
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4王娟,王桂萍,张红生,夏妍,沈振国.海州香薷金属硫蛋白(EhMT1)的原核表达和纯化及其多克隆抗体的制备[J].南京农业大学学报,2010,33(4):25-30. 被引量：3
5刘宇,寻晓红.小鼠Connexin31蛋白磷酸化定位[J].湖南理工学院学报（自然科学版）,2009,22(2):55-59.
6阮振超,朱杏丽,褚凯丽,李元春,黄燕.间隙连接蛋白Cx31.1相互作用蛋白的初步鉴定[J].生殖与避孕,2013,33(7):433-437.
7黄亮群,施小六,谭志平,房海燕,郑多,夏家辉,夏昆.Cx31相互作用蛋白:p11在酵母中表达可能存在翻译移码[J].生物化学与生物物理学报,2002,34(2):219-224.
8生物技术[J].科技开发动态,2005(4):31-32.
9江宁,孙万儒.短梗霉多糖超滤器的计算[J].生物工程学报,1991,7(3):252-256. 被引量：1
10寻晓红,刘宇,聂东宋.DSCR1抗体制备与应用[J].湖南理工学院学报（自然科学版）,2006,19(1):80-83.

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MAP法制备抗人Connexin31抗体

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