ab177171 staining Histone H2B (crotonyl K12) in HeLa cells. The cells were fixed with 100% methanol (5min) and then permeabilised using 0.1% PBS-Triton. Cells were then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab177171 at 5μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Rabbit secondary (ab150081) at 1/1000 dilution (shown in green). AlexaFluor®594 Mouse monoclonal [DM1A] to alpha Tubulin (ab195889) - Microtubule Marker was used at a 1/200 dilution and incubated for 1h with the cells, to label Microtubules (shown in Red). The nuclear counter stain is DAPI (blue), which was added to the secondary antibody mixture.