Figure 1. Western blot analysis of STUB1 using anti-STUB1 antibody (M01236). 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 50ug of sample under reducing conditions. Lane 1: human HEK293 whole cell lysates, Lane 2: monkey COS-7 whole cell lysates, Lane 3: human CACO-2 whole cell lysates. Lane 4: human THP-1 whole cell lysates, Lane 5: human PC-3 whole cell lysates, Lane 6: human U87 whole cell lysates, Lane 7: rat heart tissue lysates, Lane 8: rat brain tissue lysates, Lane 9: mouse brain tissue lysates, Lane 10: mouse HEPA1-6 whole cell lysates. 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 mouse anti-STUB1 antigen affinity purified monoclonal antibody (Catalog # M01236) 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-mouse 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 # EK1001) with Tanon 5200 system. A specific band was detected for STUB1 at approximately 35KD. The expected band size for STUB1 is at 35KD.