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| Gas8蛋白的抗体制备及小鼠组织表达图谱分析 | |
| 引用本文： | 付宁,赵莉霞,朱恒奇,周晓巍,马学恩,黄培堂.Gas8蛋白的抗体制备及小鼠组织表达图谱分析[J].生物技术通讯,2009,20(3):343-345. |
| 作者姓名： | 付宁 赵莉霞 朱恒奇 周晓巍 马学恩 黄培堂 |
| 作者单位： | 1. 军事医学科学院,生物工程研究所,北京,100071;内蒙古农业大学,动物科学与医学学院,内蒙古,呼和浩特,010019 2. 军事医学科学院,生物工程研究所,北京,100071 3. 内蒙古农业大学,动物科学与医学学院,内蒙古,呼和浩特,010019 |
| 摘 要： | 目的：制备Gas8抗体,并检测小鼠各组织中Gas8表达谱。方法：在大肠杆菌BL21（DE3）中表达His标签标记的Gas8融合蛋白,以纯化的Gas8重组融合蛋白为抗原制备兔Gas8多抗血清,进一步利用亲和纯化方法制备Gas8多克隆抗体;为确定Gas8抗体的特异性,通过免疫印迹方法检测瞬时表达的Flag-Gas8融合蛋白,同时以抗Flag单克隆抗体作为对照;利用制备的抗体通过Westernblot检测Gas8在小鼠各组织中的表达谱。结果：与Gas8特异性结合的抗体能够通过免疫法获得;利用制备的抗体检测到Gas8在小鼠心、肺、肾、脑、肝、脾、肌肉、睾丸各组织中均表达。结论：获得了特异性抗Gas8抗体,用该抗体可以检测到小鼠组织中Gas8的表达。 |
| 关 键 词： | Gas8 抗体 亲和纯化 组织表达谱 |
Preparation of Gas8 Antibody and Analysis of Gas8 Expression in Mouse Tissues |
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| FU Ning,ZHAO Li-Xia,ZHU Heng-Qi,ZHOU Xiao-Wei,MA Xue-En,HUANG Pei-Tang.Preparation of Gas8 Antibody and Analysis of Gas8 Expression in Mouse Tissues[J].Letters in Biotechnology,2009,20(3):343-345. | |
| Authors: | FU Ning ZHAO Li-Xia ZHU Heng-Qi ZHOU Xiao-Wei MA Xue-En HUANG Pei-Tang |
| Institution: | 1. Beijing Institute of Biotechnology, Beijing 100071; 2. Inner Mongolia Agriculture University, Huhehot 010019; China) |
| Abstract: | Objective： To prepare Gas8 antibody and check the expression of Gas8 protein in the tissues of the mouse. Methods： His-tagged Gas8 was expressed in E.coli BL21（DE3）. Rabbit Gas8 polyclonal antiserum was generated by injecting the purified recombinant His-tagged Gas8 inclusion body as the antigen, and further separated by affinity purification. To confirm the specificity of the Gas8 antibody, transiently expressed Flag-Gas8 fusion protein was analyzed by immunoblotting with anti-Flag monoclonal antibody as control. The produced antibody was used to check the expression of Gas8 protein in mouse tissues by Western blot. Results： Antibodies Specifically binding to Gas8 could be obtained by immunization, and expression of Gas8 was detected in heart, lung, kidney, brain, liver, spleen, muscle and testis of mouse with the generated antibody. Conclusion： The specific anti-Gas8 antibody was obtained and it could be used to detected the expression of Gas8 protein in mouse tissues. |
| Keywords: | Gas8 |
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