Western blot analysis of RENT1/hUPF1 in, Lane 1: human Hela whole cell lysates, Lane 2: human Raji whole cell lysates, Lane 3: human HepG2 whole cell lysates, Lane 4: human SK-OV-3 whole cell lysates, Lane 5: human PC-3 whole cell lysates, Lane 6: human HEK293 whole cell lysates, Lane 7: rat RH35 whole cell lysates, Lane 8: mouse HEPA1-6 whole cell lysates. 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. After Electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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 UPF1 Polyclonal Antibody (Product # PA5-80208) 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 1:5,000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit. A specific band was detected for RENT1/hUPF1 at approximately 140 kDa. The expected band size for RENT1/hUPF1 is at 124 kDa.

Immunohistochemistry analysis of UPF1 on paraffin-embedded human intestinal cancer tissue. Sample was incubated with UPF1 polyclonal antibody (Product# PA5-80208).