All lanes : Anti-Histone H2B (acetyl K16) antibody - ChIP Grade ( ab40977 ) at 1 µg/ml Lane 1 : HeLa Histone Preparation Nuclear Lysate Lane 2 : HeLa Histone Preparation Nuclear Lysate - Butyrated treated Lane 3 : HeLa Histone Preparation Nuclear Lysate with Human Histone H2B (acetyl K16) peptide (ab41879) at 1 µg/ml Lane 4 : HeLa Histone Preparation Nuclear Lysate - Butyrated treated with Human Histone H2B (acetyl K16) peptide (ab41879) at 1 µg/ml Lane 5 : HeLa Histone Preparation Nuclear Lysate with Human Histone H2B peptide ( ab35793 ) at 1 µg/ml Lane 6 : HeLa Histone Preparation Nuclear Lysate - Butyrated treated with Human Histone H2B peptide ( ab35793 ) at 1 µg/ml Lysates/proteins at 2.5 µg per lane. Secondary HRP Conjugated Goat Anti-Rabbit IgG at 1/3000 dilution Performed under reducing conditions. Predicted band size : 14 kDa Observed band size : 16 kDa ( why is the actual band size different from the predicted? )