Anti ZAP70, Picoband^TM  

Figure 1. Western blot analysis of ZAP70 using anti-ZAP70 antibody (M00754-4).
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 Jurkat whole cell lysates,
Lane 2: human CCR-CEM whole cell lysates,
Lane 3: mouse spleen tissue lysates,
Lane 4: mouse thymus tissue 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-ZAP70 antigen affinity purified monoclonal antibody (Catalog # M00754-4) 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 (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for ZAP70 at approximately 70KD. The expected band size for ZAP70 is at 70KD.