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Spirende Gær Protein ekstraktion og oprensning for Studiet af Function, Interactions og post-translationelle modifikationer
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JoVE Journal Biology
Budding Yeast Protein Extraction and Purification for the Study of Function, Interactions, and Post-translational Modifications

Spirende Gær Protein ekstraktion og oprensning for Studiet af Function, Interactions og post-translationelle modifikationer

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

Udarbejdelsen af ​​høj kvalitet gær celleekstrakter er et nødvendigt første skridt i analysen af ​​de enkelte proteiner eller hele proteomer. Her beskriver vi en hurtig, effektiv og pålidelig homogenisering protokol for spirende gærceller, der er blevet optimeret til at bevare protein funktioner, interaktioner og post-translationelle modifikationer.

Tags

Budding YeastProtein ExtractionProtein PurificationBead BeatingHomogenizationCell LysisPost-translational ModificationsProtein InteractionsAffinity PurificationSUMOSiz1Slx5

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