In vitro ubiquitination and degradation of bacteria-purified human histone H2B by the nucleoli fractions

Histones are the main components of chromatin, and the protein levels of histones significantly affect chromatin assembly. Here, we describe detailed protocols for histone H2B purification from bacteria and for the separation of nucleolar fractions and cytoplasmic and nucleoplasmic fractions. Finally, the in vitro ubiquitination and degradation of H2B by distinct cellular fractions are described. For complete details on the use and execution of this protocol, please refer to Liu et al. (2021).