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In Vitro Characterization of Histone Chaperones using Analytical, Pull-Down and Chaperoning Assays
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JoVE 신문 생화학
In Vitro Characterization of Histone Chaperones using Analytical, Pull-Down and Chaperoning Assays
DOI:

08:16 min

December 29, 2021

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Chapters

  • 00:05Introduction
  • 01:04Competitive Pull-Down Assay
  • 02:06Analytical Ultracentrifugation-Sedimentation Velocity (AUC-SV) Experiments
  • 04:29Plasmid Supercoiling Assay to Confirm Histone Chaperoning Function
  • 06:11Results: Characterization of Histone Chaperones Using Analytical, Pull-Down, and Chaperoning Assays
  • 07:41Conclusion

Summary

자동 번역

This protocol describes a battery of methods that includes analytical size-exclusion chromatography to study histone chaperone oligomerization and stability, pull-down assay to unravel histone chaperone-histone interactions, AUC to analyze the stoichiometry of the protein complexes, and histone chaperoning assay to functionally characterize a putative histone chaperone in vitro.

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