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Simultaneous Measurement of HDAC1 and HDAC6 Activity in HeLa Cells Using UHPLC-MS
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JoVE Journal Chimica
Simultaneous Measurement of HDAC1 and HDAC6 Activity in HeLa Cells Using UHPLC-MS

Simultaneous Measurement of HDAC1 and HDAC6 Activity in HeLa Cells Using UHPLC-MS

DOI:
10.3791/55878-v

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09:20 min

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August 10, 2017

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, , ,
1Section des Sciences Pharmaceutiques,Universités des Geneva–Lausanne (EPGL)

Capitoli

  • 00:05Titolo
  • 01:20Cell Culture
  • 03:00Cell Treatment with Cell Compounds and HDAC Substrates, Cell Lysis, and Sample Preparation for UHPLC-ESI-MS/MS
  • 05:14UHPLC/MS-MS Analysis and Data Analysis
  • 06:14Results: Deacetylation of Specific Substrates by Endogenous HDACs and Potency of Standard HDAC Inhibitors
  • 08:18Conclusion

Summary

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Traduzione automatica

The present method serves to identify isoform-specific inhibitors of histone deacetylases (HDAC) in HeLa cells by the UHPLC-MS analysis of multiple substrates. This is an antibody-free method developed to reflect HDAC1 and HDAC6 activity in the living cell environment, in contrast to single-isoform cell-free assays.

Tags

HDAC1HDAC6HeLa CellsUHPLC-MSHDAC InhibitorSelectivityDrug DiscoverySimultaneous MeasurementCell-based AssayAcetylationTrichostatin AMS275Tubastatin A

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