|本期目录/Table of Contents|

[1]朱雄伟,张佑红,徐智鹏,等.自噬基因融合蛋白重组多克隆抗体的纯化与检测[J].武汉工程大学学报,2013,(02):37-41.[doi:103969/jissn16742869201302008]
　ZHU Xiong wei,ZHANG You hong,XU Zhi peng,et al.Purification and detection of autophagyrelated gene fusion protein restructured polyclonal antibody[J].Journal of Wuhan Institute of Technology,2013,(02):37-41.[doi:103969/jissn16742869201302008]
点击复制

自噬基因融合蛋白重组多克隆抗体的纯化与检测(/HTML)
分享到：

《武汉工程大学学报》[ISSN:1674-2869/CN:42-1779/TQ]

卷:
期数:
2013年02期
页码:
37-41
栏目:
化学与化学工程
出版日期:
2013-02-28

文章信息/Info

Title:
Purification and detection of autophagyrelated gene fusion protein restructured polyclonal antibody
文章编号:
16742869(2013)02003705
作者:
朱雄伟; 张佑红; 徐智鹏; 苏腾甲; 熊瑶; 翟莉莉
武汉工程大学化工与制药学院，绿色化工过程省部共建教育部重点实验室，湖北 武汉 430074
Author(s):
ZHU Xiongwei; ZHANG Youhong; XU Zhipeng; SU Tengjia; XIONG Yao; ZHAI Lili
Key Laboratory for Green Chemical Process of Ministry of Education, School of Chemical Engineering and Pharmacy, Wuhan Institute of Technology, Wuhan 430074, China
关键词:
. 多克隆抗体; 免疫亲和纯化; 酶联免疫吸附法; 蛋白印迹; 聚丙烯酰胺凝胶电泳
Keywords:
polyclonal antibody; immune affinity purification; enzyme linked immunosorbent assay; Western blotting; polyacrylamide gel electrophoresis
分类号:
R392.1
DOI:
103969/jissn16742869201302008
文献标志码:
A
摘要:
为了提纯并鉴定自噬基因融合蛋白重组多克隆抗体，探讨其在部分组织中的免疫定位.首先合成带谷胱甘肽转移酶标签的自噬基因融合蛋白，免疫新西兰大白兔，制备多克隆抗体，然后用聚丙烯酰胺凝胶电泳检测抗体浓度，最后用免疫亲和纯化方法提纯抗血清中的多克隆抗体，酶联免疫吸附法检测抗体效价，蛋白印迹检测抗体特异性及在组织中的表达情况.结果表明：制备得到自噬基因融合蛋白重组多克隆抗体经聚丙烯酰胺凝胶电泳检测其质量浓度为每10 μL 含5 μg，用酶联免疫吸附法显示此抗体效价及特异性较高，蛋白印迹显示蛋白大小为55 kD左右
Abstract:
The aim of the study is to purify and identify the autophagyrelated gene fusion protein restructured polyclonal antibody, and to detect the immune localization in tissues. Firstly, the autophagyrelated gene fusion protein restructured polyclonal antibody with glutathioneStransferase tag was synthesized, the New Zealand white rabbits was immuned and the polyclonal antibody was prepared. Secondly, the antibody concentration was detected by polyacrylamide gel electrophoresis. Finally, the antibody was purified in the antiserum by using immunoaffinity purification, antibody titer was detected by enzyme linked immunosorbent assay methods, the antibody specificity and the organization expression were tested by Western blotting.The result shows that autophagyrelated gene fusion protein restructured polyclonal antibody concentration is 10 μL per 5 μg, which is detected by polyacrylamide gel electrophoresis; the antibody is proved to have higher titer and specificity by enzyme linked immunosorbent assay; the molecular weight of the protein is about 55 kD from Western blotting results.

参考文献/References:

［1］周建光,杨梅. 免疫学技术的发展及临床应用\[J\].医疗装备,2011,24(7): 4951.［2］孔维,杨文辉. 单克隆抗休及其应用的研究进展\[J\]畜牧兽医科技信息,2009(1): 911.［3］Ashford T P, Porter K R. Cytoplasmic components in hepatic cell lysosomes\[J\].The Journal of cell biology,1962(1):198202.［4］谭荣荣,刘明炎,龚自明. 茶尺蠖核型多角体病毒多克隆抗体的制备及检测应用\[J\]. 湖北农业科学,2012,51(2): 298301.［5］张花美,李因来,潘熙萍. 抗麦草畏多克隆抗体v 的制备及其间接竞争ELISA检测方法的建立\[J\]. 浙江大学学报:农业与生命科学版,2012,38(2): 153158.［6］Susumu H T, Suzuki K, Akahori Y. Antibodydependent cellmediated cytotoxicity is induced by a singlechain Fvprotein III fusion in the presence of a rabbit antiprotein III polyclonalantibody\[J\]. Immunology Letters,2011,136(1):4448.［7］Horiguchi T, Urushitani H, Ohta Y. Establishment of a polyclonal antibody against the retinoid X receptor of the rock shell Thais clavigera and its application to rock shell tissues for imposex research\[J\]. Ecotoxicology,2010,19(3):571576.［8］Xu M Y, Wang S D. Prokaryotic expression of pathogenesis related protein 1 gene from Nicotiana benthamiana: antifungal activity and preparation of its polyclonal antibody\[J\]. Biotechnology Letters,2012,34(5):919924.［9］Anderson G J, Cipolla C M, Kennedy R T. Western Blotting Using Capillary Electrophoresis\[J\]. Analytical chemistry, 2011, 83 (4):13501355.［10］Welinder C, Ekblad L.Coomassie Staining as Loading Control in Western Blot Analysis\[J\].Journal of proteome research,2011,10(3): 14161419.［11］Rath A, Glibowicka M, Nadeau V G. Detergent binding explains anomalous SDSPAGE migration of membrane proteins\[J\]. Proceedings of the National Academy of Sciences of the United States of America, 2009,106(6):17601765.［12］Hans J C, Wessels R O. LCMS/MS as an alternative for SDSPAGE in blue native analysis of protein complexes\[J\]. Cell & Molecular Biology, 2009,9(17):42214228.［13］Riechers A, Schmidt F, Stadlbauer S. Detection of Protein Phosphorylation on SDSPAGE Using Probes with a PhosphateSensitive Emission Response\[J\]. Bioconjugate Chemistry, 2009, 20 (4):804807.

相似文献/References:

备注/Memo

备注/Memo:
收稿日期：20120906作者简介：朱雄伟（1973），男，湖北咸宁人，讲师，博士研究生.研究方向：生物化学与生物学.
更新日期/Last Update: 2013-04-16