14-3-3 zeta/delta/YWHAZ/14 Mouse Monoclonal Antibody

Western blot analysis of 14-3-3 zeta/delta using anti-14-3-3 zeta/delta 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 50 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates; Lane 2: human A549 whole cell lysates; Lane 3: monkey COS-7 whole cell lysates; Lane 4: human Raji whole cell lysates; Lane 5: huamn Caco-2 whole cell lysates; Lane 6: huamn Jurkat whole cell lysates; Lane 7: mouse brain tissue lysates; Lane 8: rat brain tissue 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-14-3-3 zeta/delta 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 14-3-3 zeta/delta at approximately 28 KD. The expected band size for 14-3-3 zeta/delta is at 28 KD.