Western blot analysis of APE1 using anti-APE1 antibody (M00627). 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 50ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human MCF-7 whole cell lysates, Lane 3: human COLO-320 whole cell lysates, Lane 4: human HepG2 whole cell lysates, Lane 5: Rabbit IgG, Lane 6: Marker 1113, Lane 7: human A549 whole cell lysates, Lane 8: human PANC-1 whole cell lysates, Lane 9: human 22RV1 whole cell lysates, Lane 10: human MDA-MB-453 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-APE1 antigen affinity purified monoclonal antibody (Catalog # M00627) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system.