All lanes : Anti-Histone H2A.X (phospho Y142) antibody (ab94602) at 1 µg/mlLane 1 : Calf Thymus Histone Preparation Nuclear LysateLane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H2A.X (phospho Y142) peptide (ab113997) at 1 µg/mlLane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H2A.X peptide (ab114003) at 1 µg/mlLysates/proteins at 20 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ICC/IF image of ab94602 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab94602, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.