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In Vitro Characterization of Histone Chaperones using Analytical, Pull-Down and Chaperoning Assays
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Capitoli
- 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
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.
