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Deacetylation Assays to Unravel the Interplay between Sirtuins (SIRT2) and Specific Protein-substrates
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JoVE Journal Chemistry
Deacetylation Assays to Unravel the Interplay between Sirtuins (SIRT2) and Specific Protein-substrates

Deacetylation Assays to Unravel the Interplay between Sirtuins (SIRT2) and Specific Protein-substrates

DOI:
10.3791/53563-v

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14:32 min

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February 27, 2016

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, , ,
1Laboratory for Molecular Cancer Biology, Department of Radiation Oncology, Robert H. Lurie Comprehensive Cancer Center,Feinberg School of Medicine, Northwestern University, 2Department of Radiation Oncology, Robert H. Lurie Comprehensive Cancer Center,Feinberg School of Medicine, Northwestern University

Chapters

  • 00:05Title
  • 01:06Purification of SIRT2
  • 06:39Purification of Acetylated Protein-substrate
  • 09:33In Vitro Deacetylation Reaction
  • 11:11Results: SIRT2 Deacetylates of Protein-substrate
  • 13:00Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes the required steps to execute in vitro and in vivo deacetylation assays in order to establish the role of proteins as specific deacetylation substrates for sirtuins and further study the role of reversible - lysine acetylation as a post-translational modification.

Tags

Keywords: Deacetylation AssaySirtuinsSIRT2Protein SubstratesIn VitroIn VivoHEK 293T CellsLentivirusTransfectionOverexpressionPuromycin Selection

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