武汉工程大学学报2013，Vol.35Issue(2)：37-41,5.DOI:10.3969/j.issn.1674-2869.2013.02.008
自噬基因融合蛋白重组多克隆抗体的纯化与检测
Purification and detection of autophagy-related gene fusion protein restructured polyclonal antibody
朱雄伟 1张佑红 1徐智鹏 1苏腾甲 1熊瑶 1翟莉莉1
作者信息
摘要
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.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Key words
polyclonal antibody/ immune affinity purification/ enzyme linked immunosorbent assay/ Western blotting/polyacrylamide gel electrophoresis分类
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朱雄伟,张佑红,徐智鹏,苏腾甲,熊瑶,翟莉莉..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,2013,35(2):37-41,5.
