Anti-14-3-3 zeta/delta/YWHAZ Antibody Picoband™ (monoclonal, 6G5)

Figure 1. Western blot analysis of 14-3-3 zeta/delta using anti-14-3-3 zeta/delta antibody (M01141).
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 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 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-14-3-3 zeta/delta antigen affinity purified monoclonal antibody (Catalog # M01141) 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 14-3-3 zeta/delta at approximately 28KD. The expected band size for 14-3-3 zeta/delta is at 28KD.

Figure 2. Flow Cytometry analysis of PC-3 cells using anti-14-3-3 zeta/delta antibody (M01141).
Overlay histogram showing PC-3 cells stained with M01141 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-14-3-3 zeta/delta Antibody (M01141,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 3. Flow Cytometry analysis of SiHa cells using anti-14-3-3 zeta/delta antibody (M01141).
Overlay histogram showing SiHa cells stained with M01141 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-14-3-3 zeta/delta Antibody (M01141,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.