University of Vermont College of Medicine 1 article published in JoVE Biochemistry In Vitro Characterization of Histone Chaperones using Analytical, Pull-Down and Chaperoning Assays Ruchir C. Bobde*1,2, Ketul Saharan*1,2, Somanath Baral1,3, Surajit Gandhi1,2, Archana Samal1,2, Rajivgandhi Sundaram1, Ashish Kumar1,4, Ajit K. Singh1,5, Aritreyee Datta1, Dileep Vasudevan1 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 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.
