Western blot analysis of nmt55 p54nrb using anti-nmt55 p54nrb 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 50ug of sample under reducing conditions. Lane 1: human HELA whole cell lysate, Lane 2: human Placent tissue lysate, Lane 3: human MCF-7 whole cell lysate, Lane 4: human A549 whole cell lysate, Lane 5: human SW620 whole cell lysate, Lane 6: human PANC-1 whole cell lysate, Lane 7: human U20S whole cell lysate, Lane 8: human K562 whole cell lysate. 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-nmt55 p54nrb 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.