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Assays for Validating Histone Acetyltransferase Inhibitors
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JoVE Journal Ricerca sul cancro
Assays for Validating Histone Acetyltransferase Inhibitors

Assays for Validating Histone Acetyltransferase Inhibitors

DOI:
10.3791/61289-v

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09:11 min

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August 06, 2020

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,
1Department of Anatomy and Cell Biology, and UF Health Cancer Center,University of Florida College of Medicine

Capitoli

  • 00:04Introduction
  • 00:39In Vitro HAT Assay
  • 01:37ChHAI Assay
  • 03:11Chromatin Immunoprecipitation (ChIP)
  • 05:48Results: Inhibitors of Histone Acetyltransferases
  • 08:19Conclusion

Summary

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Inhibitors of histone acetyltransferases (HATs, also known as lysine acetyltransferases), such as CBP/p300, are potential therapeutics for treating cancer. However, rigorous methods for validating these inhibitors are needed. Three in vitro methods for validation include HAT assays with recombinant acetyltransferases, immunoblotting for histone acetylation in cell culture, and ChIP-qPCR.

Tags

Histone AcetyltransferaseHAT InhibitorEpigenetic RegulationEnzymatic ReactionHistone AcetylationSDS-PAGEImmunoblottingMCF-7 CellsMS275A-485Passive Lysis BufferChromatin Immunoprecipitation

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