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JoVE Journal Biochemistry
In Vitro Characterization of Histone Chaperones using Analytical, Pull-Down and Chaperoning Assays
 

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

Article DOI: 10.3791/63218-v • 08:16 min • December 29th, 2021
December 29th, 2021 •
*1,2, *1,2, 1,3, 1,2, 1,2, 1, 1,4, 1,5, 1, 1
1Institute of Life Sciences, 2Regional Centre for Biotechnology, 3School of Biotechnology, KIIT University, 4Department of Molecular Biophysics and Biochemistry, Yale University, 5Department of Pharmacology, University of Vermont College of Medicine
* These authors contributed equally

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Resumo
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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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Keywords: Histone Chaperones Pull-down Assays Analytical Size Exclusion Chromatography Analytical Ultra-centrifugation Histone Chaperoning Assay Chromatin Research H2A/H2B Dimer H3/H4 Tetramer SDS-PAGE Coomassie Brilliant Blue R250 Dialysis Buffer Analytical Ultra Centrifuge
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