OPA1

Figure 1. Western blot analysis of OPA1 using anti-OPA1 antibody Electrophoresis was performed on a 5-20% SDS-PAGE gel at70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions. Lane 1: human MCF-7 whole cell lysates, Lane 2: human PANC-1 whole cell lysates, Lane 3: human PC-3 whole cell lysates, Lane 4: human T47D 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 rabbit anti- OPA1 antigen affinity purified monoclonal antibodyat 0.5 mug/mL 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 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for OPA1 at approximately 80-100KD. The expected band size for OPA1 is at 80-100KD.