Western blot analysis of mtTFA in, Lane 1: Caco-2 whole cell lysates, Lane 2: SW620 whole cell lysates, Lane 3: PANC-1 whole cell lysates, Lane 4: rat brain tissue lysates, Lane 5: mouse brain tissue lysates. 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 50 µg of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. The membrane was blocked with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with TFAM Polyclonal Antibody (Product # PA5-80107) 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 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit. A specific band was detected for mtTFA at approximately 24KD. The expected band size for mtTFA is at 24KD.