Western blot analysis of NOP10 using anti-NOP10 antibody (M07394).
Electrophoresis was performed on a 12% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human A549 whole cell lysates,
Lane 2: human PC-3 whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human RT4 whole cell lysates,
Lane 5: human HEL whole cell lysates,
Lane 6: human THP-1 whole cell lysates,
Lane 7: human 293T whole cell lysates,
Lane 8: human MCF-7 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 rabbit anti-NOP10 antigen affinity purified monoclonal antibody (M07394) at 1:500 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 at a dilution of 1:500 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 NOP10 at approximately 10 kDa. The expected band size for NOP10 is at 8 kDa.