Profiling histone post-translational modifications (PTMs) in clinical samples holds great potential for the identification of epigenetic biomarkers and the discovery of novel epigenetic targets. We have recently developed a battery of mass spectrometry (MS)-based approaches to analyze histone PTMs in different types of primary samples. However, most of these protocols rely on SDS-PAGE separation following histone enrichment in order to eliminate detergents and isolate histones from other proteins present in the sample. As a consequence, many proteolytic enzymes, whose performance is poor in gel, cannot be used, limiting the digestions options for clinical samples, and hence the modification coverage. In this study, we used a simple procedure involving acetone protein precipitation followed by histone enrichment through a C18 StageTip column to obtain histone preparations suitable for various in solution digestion protocols.