Figure 1. Western blot analysis of ANGPTL4 using ANGPTL4. Electrophoresis was performed on a 5-20% SDS- PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. lane 1: recombinant human ANGPTL4 protein 1ng After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with ANGPTL4 at 0.5ug/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat 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 with Tanon 5200 system. A specific band was detected for ANGPTL4 at approximately 55kD. The expected band size for ANGPTL4 is at 44kD.