Western blot analysis of Acetyl-Histone H4 (Lys12) was performed by loading 20 µg of HeLa soluble nuclear and HeLa chromatin bound cell lysate fractions prepared using a Subcellular Protein Fractionation Kit (Product # 78840) in addition to 10 ng of recombinant H4 (Lys12), H4 (Lys8), and H4 (Lys5) protein and 10 µL PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288) and blocked with 5% Milk/TBST for at least 1 hour at room temperature. Acetyl-Histone H4 (Lys12) was detected using a Acetyl-Histone H4 (Lys12) mouse monoclonal antibody, Product # MA3-058, at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform, followed by a Superclonal Goat anti-Mouse IgG-HRP secondary antibody (Product # A28177) at a dilution of 1:2,000 for at least 1 hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076) and the myECL Imager (Product # 62236).