All lanes : Anti-Histone H2B (acetyl K20) antibody (ab175361) at 1 µg/mlLane 1 : HeLa Sodium Butyrate-treated 0uM Control (Cell acid extract)Lane 2 : HeLa Sodium Butyrate-treated 10uM Control (Cell acid extract)Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 4 : Calf Thymus Histone Preparation Nuclear LysateLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (Alexa Fluor® 790) at 1/10000 dilutionPerformed under reducing conditions.
ab175361 staining Histone H2B type 2-E (Acetylation K20) in HeLa cells. The cells were fixed with 100% methanol (5 min) 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 overnight at +4°C with ab175361 at 1ug/ml and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594) at 2µg/ml (Shown in red). This was followed by an incubation at room temperature for 1h with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).