Figure 1. Western blot analysis of HistoneH2A.X using anti-HistoneH2A.X antibody (MP00241). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human Jurkat whole cell lysates, Lane 2: human U20S whole cell lysates, Lane 3: human PC-3 whole cell lysates, Lane 4: human 293T whole cell lysates, Lane 5: rat PC-12 whole cell lysates, Lane 6: rat C6 whole cell lysates, Lane 7: mouse NIH/3T3 whole cell lysates, Lane 8: mouse Neuro-2a whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-HistoneH2A.X antigen affinity purified monoclonal antibody (Catalog # MP00241) at 1:5000 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:1000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for HistoneH2A.X at approximately 15 kDa. The expected band size for HistoneH2A.X is at 15 kDa.