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从Cryomilled哺乳动物细胞中的蛋白质复合物亲和捕获

JoVE 杂志
生物化学

This content is Free Access.

JoVE 杂志生物化学

Protein Complex Affinity Capture from Cryomilled Mammalian Cells

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从Cryomilled哺乳动物细胞中的蛋白质复合物亲和捕获

DOI:

10.3791/54518-v

•

10:37 min

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December 09, 2016

•

John LaCava², Hua Jiang, Michael P. Rout

¹Laboratory of Cellular and Structural Biology,The Rockefeller University, ²Institute for Systems Genetics, Department of Biochemistry and Molecular Pharmacology,New York University School of Medicine

Chapters

00:05Title
00:30Cell Harvesting and Freezing
03:09Cryogenic Disruption of Frozen Cells
04:50Affinity Capture of Protein Complexes from Cell Extracts
06:02Affinity Capture
07:41Results: Comparing Some Different Aspects of Affinity Capture Optimization
09:38Conclusion

Summary

自动翻译

这里，我们描述协议破坏由固态研磨哺乳动物细胞在低温下，产生从所得细胞粉细胞提取物，并且在抗体偶联微米级的顺磁珠亲和捕获隔离感兴趣的蛋白质复合物。

Tags

Protein Complex Affinity Capture Cryomilled Mammalian Cells Cell Biology Molecular Biology Protein Interactions PBS Cell Pellet Liquid Nitrogen Cell Lysis

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