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Budding Yeast Protein Extraction and Purification for the Study of Function, Interactions, and Post-translational Modifications
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JoVE Journal Biology
Budding Yeast Protein Extraction and Purification for the Study of Function, Interactions, and Post-translational Modifications

Budding Yeast Protein Extraction and Purification for the Study of Function, Interactions, and Post-translational Modifications

DOI:
10.3791/50921-v

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09:22 min

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October 30, 2013

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1Integrated Science Center, Biology Department,The College of William & Mary

Chapters

  • 00:05Title
  • 01:19Purification of 6xHIS-tagged Proteins Expressed in Budding Yeast Cells Under Native Conditions
  • 03:01Homogenization of Yeast Cells and Extraction of Proteins
  • 04:06Batch Purification of Proteins from Yeast Cell Homogenates
  • 06:23Representative Yeast Extraction and Purification Results
  • 08:57Conclusion

Summary

Automatic Translation

Automatic Translation

The preparation of high quality yeast cell extracts is a necessary first step in the analysis of individual proteins or entire proteomes. Here we describe a fast, efficient, and reliable homogenization protocol for budding yeast cells that has been optimized to preserve protein functions, interactions, and post-translational modifications.

Tags

Keywords: Budding YeastProtein ExtractionProtein PurificationBead BeatingHomogenizationCell LysisPost-translational ModificationsProtein InteractionsAffinity PurificationSUMOSiz1Slx5

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