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DOI :
10.3791/4057-v
November 12th, 2012
Chapters
0:05
Title
1:47
Construction of Gene-specific SPA-tagging in E. Coli DY330 Strain
3:54
Culturing and Sonication
5:26
Affinity Purification
7:53
Proteolysis and Sample Preparation for Mass Spectrometry
9:38
Protein Identification by LTQ Orbitap Velos Mass Spectrometer
11:02
Results
14:27
Conclusion
亲和纯化标签蛋白结合质谱(APMS)是一个功能强大的蛋白质相互作用网络系统的映射方法和调查的生物过程的机理的基础。在这里,我们描述了一个优化的顺序肽亲和力(SPA)APMS程序开发的细菌大肠杆菌,可用于分离和表征到接近均匀性,甚至从每个细胞的拷贝数低的稳定的多蛋白复合物。
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