Figure 1. Western blot analysis of MTCO1 using anti-MTCO1 antibody (PA1317-1). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The samples were loaded under reducing conditions. Lane 1: human HeLa mitochondria lysates at 20ug, Lane 2: human HeLa whole cell lysates at 20ug, Lane 3: human Caco-2 whole cell lysates at 50ug, Lane 4: rat heart tissue lysates at 50ug. Lane 5: mouse heart tissue lysates at 50ug. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 60 minutes. Blocked the membrane with 5% Non-fat Milk in TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-MTCO1 antigen affinity purified polyclonal antibody (Catalog # PA1317-1) 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-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 (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for MTCO1 at approximately 37KD. The expected band size for MTCO1 is at 57KD.