Figure 1. Western blot analysis of CARD12 using anti-CARD12 antibody (PB9673). 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 40 ug of sample under reducing conditions. Lane 1: 22RV1 Whole Cell Lysate, Lane 2: RH35 Whole Cell Lysate, Lane 3: NIH3T3 Whole Cell Lysate. 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 rabbit anti-CARD12 antigen affinity purified polyclonal antibody (Catalog # PB9673) 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 1:5000 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 CARD12 at approximately 116 kDa. The expected band size for CARD12 is at 116 kDa.