Figure 1. Western blot analysis of EIF3E using anti-EIF3E antibody (MBS1756546).

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 Raji whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: rat thymus tissue lysates,
Lane 4: mouse thymus tissue lysates,
Lane 5: mouse ANA-1 whole cell lysates,
Lane 6: mouse SP2/0 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-EIF3E antigen affinity purified monoclonal antibody (#MBS1756546) at 0.5 ug/mL overnight at 4 degree 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 EIF3E at approximately 52 kDa. The expected band size for EIF3E is at 52 kDa.