Figure 1. Western blot analysis of ATP5F1,2,3/ATP5MC1,2,3 using anti-ATP5F1,2,3/ATP5MC1,2,3 antibody (M09735). 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 HL-60 whole cell lysates, Lane 2: monkey COS-7 whole cell lysates, Lane 3: rat kidney tissue lysates, Lane 4: rat heart tissue lysates, Lane 5: mouse heart 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 mouse anti-ATP5F1,2,3/ATP5MC1,2,3 antigen affinity purified monoclonal antibody (Catalog # M09735) at 0.5 microg/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. A specific band was detected for ATP5F1,2,3/ATP5MC1,2,3 at approximately 10-14KD. The expected band size for ATP5F1,2,3/ATP5MC1,2,3 is at 10-14KD.