Western blot analysis of ERLIN2 using ERLIN2 antibody (Product # MA5-49296). 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 HepG2 whole cell lysates; Lane 2: human T47D whole cell lysates; Lane 3: human HCCT tissue lysates; Lane 4: human HCCP tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-ERLIN2 antigen affinity purified monoclonal antibody (Product # MA5-49296) 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-mouse IgG-HRP secondary antibody at a dilution of 1:10,000 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 ERLIN2 at approximately 43 kDa. The expected band size for ERLIN2 is at 38 kDa.