Western blot analysis of IDO2 in, Lane 1: human A549 whole cell lysates (positive control), Lane 2: human placenta tissue lysates (positive control), Lane 3: human Caco-2 whole cell lysates (positive control), Lane 4: human PC-3 whole cell lysates (negative control). 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 IDO2 Polyclonal Antibody (Product # PA5-79438) 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 IDO2 at approximately 47KD. The expected band size for IDO2 is at 47KD.