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Massespektrometrianalyse for å identifisere Ubiquitylation av EYFP-merket CENP-A (EYFP-CENP-A)
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Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A (EYFP-CENP-A)

Massespektrometrianalyse for å identifisere Ubiquitylation av EYFP-merket CENP-A (EYFP-CENP-A)

DOI:
10.3791/61138-v

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09:02 min

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June 10, 2020

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, , , , , , ,
1MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center,Nanjing University, 2Jiangsu Key Laboratory of Molecular Medicine,Medical School of Nanjing University, 3Greehey Children’s Cancer Institute, Department of Molecular Medicine,UT Health Science Center San Antonio

Chapters

  • 00:04Introduction
  • 00:44In Vivo Ubiquitylation Assays Using pQCXIP-EYFP-CENP-A
  • 02:19Mass Spectrometry to Identify the Ubiquitylation Site of the EYFP-CENP-A K124R Mutant
  • 04:55Data Analysis
  • 06:22Results: EYFP-CENP-A K124 Mutant is Ubiquitylated and Interacts with HJURP
  • 08:12Conclusion

Summary

Automatic Translation

Automatic Translation

CENP-A ubiquitylation er et viktig krav for CENP-A deponering ved centromere, arvet gjennom dimerisering mellom celledeling, og uunnværlig for celle levedyktighet. Her beskriver vi massespektrometrianalyse for å identifisere ubiquitylation av EYFP-merket CENP-A (EYFP-CENP-A) protein.

Tags

Mass SpectrometryUbiquitylationEYFP-CENP-AProtein IdentificationImmunoprecipitationGFP AntibodyLysate PreparationGel ElectrophoresisCoomassie Blue StainingProtein DigestionMass Spectrometry Analysis

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