PDE4B Antibody / Phosphodiesterase 4B (FY12523)

Western blot analysis of PDE4B using anti-PDE4B antibody. Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human whole cell lysates, Lane 2: human HepG2 whole cell lysates, Lane 3: human U251 whole cell lysates, Lane 4: mouse brain tissue lysates. 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-PDE4B antibody at 0.5 ug/ml overnight at 4oC, 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 was developed using an ECL Plus Western Blotting Substrate. PDE4B (~83 kDa predicted) was detected at 95-97 kDa, consistent with expression of the longer PDE4B5 isoform and phosphorylation-dependent mobility shift characteristic of PDE4 family enzymes.