Western blot analysis of MIEF1 using MIEF1 antibody (Product # PA5-143958). 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 µg of sample under reducing conditions. Lane 1: human HT1080 whole cell lysates; Lane 2: human Jurkat whole cell lysates; Lane 3: human K562 whole cell lysates; Lane 4: human HL-60 whole cell lysates; Lane 5: human HEK293 whole cell lysates; Lane 6: human U-937 whole cell lysates; Lane 7: human HepG2 whole cell lysates; Lane 8: rat heart tissue lysates; Lane 9: rat kidney tissue lysates; Lane 10: mouse kidney tissue 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-MIEF1 antigen affinity purified polyclonal antibody (Product # PA5-143958) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5,000 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 MIEF1 at approximately 51KD. The expected band size for MIEF1 is at 51KD.