ab176430 staining Histone H2B (acetyl K120) in HeLa cells. The cells were fixed with 4% PFA (10min), permabilised with 0.1%Triton (5min) and 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 ab176430 at 1μg/ml and ab195889 at 2µg/ml (shown in pseudo color red) overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Rabbit secondary (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
All lanes : Anti-Histone H2B (acetyl K120) antibody (ab176430) at 1 µg/mlLane 1 : HeLa Sodium Butyrate treated 0uM Control (Cell acid extract) at 10 µgLane 2 : HeLa Sodium Butyrate treated 10uM (Cell acid extract) at 10 µgLane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µgLane 4 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µgSecondaryGoat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) at 1/10000 dilutionPerformed under reducing conditions.