Ikaros/IKZF1 Antibody (monoclonal, 5F12H7)

Western blot analysis of Ikaros/IKZF1 using anti-Ikaros/IKZF1 antibody. 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 ug of sample under reducing conditions. Lane 1: human Daudi whole cell lysates, Lane 2: human Jurkat whole cell lysates, Lane 3: human Raji whole cell lysates, Lane 4: human K562 whole cell 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 mouse anti-Ikaros/IKZF1 antigen affinity purified monoclonal antibody 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-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 with Tanon 5200 system. A specific band was detected for Ikaros/IKZF1 at approximately 55-65 kDa. The expected band size for Ikaros/IKZF1 is at 58 kDa.