Figure 1. Western blot analysis of Emerin using anti-Emerin antibody (M00714).
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 50ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human placenta tissue lysates,
Lane 3: human Caco-2 whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: Rabbit IgG,
Lane 6: Marker 1113,
Lane 7: human Jurkat whole cell lysates.
Lane 8: human MDA-MB-453 whole cell lysates,
Lane 9: human SK-OV-3 whole cell lysates,
Lane 10: human SW620 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-Emerin antigen affinity purified monoclonal antibody 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 Biotin Conjugated goat anti-mouse IgG 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.