Western blot analysis of IL1B using anti-IL1B antibody (PA1533).
Electrophoresis was performed on a 13% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours.
Lane 1: recombinant rat IL1B protein 10 ng.
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-IL1B antigen affinity purified polyclonal antibody (PA1533) 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 (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for IL1B at approximately 17 kDa.