Anti-PANX3 Antibody Picoband®

Western blot analysis of PANX3 using anti-PANX3 antibody (A11546-1).
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 30 ug of sample under reducing conditions.
Lane 1: human MCF-7 whole cell lysates,
Lane 2: human A431 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-PANX3 antigen affinity purified polyclonal antibody (Catalog # A11546-1) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for PANX3 at approximately 45 kDa. The expected band size for PANX3 is at 45 kDa.

Flow Cytometry analysis of MCF-7 cells using anti-PANX3 antibody (A11546-1).
Overlay histogram showing MCF-7 cells stained with A11546-1 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-PANX3 Antibody (A11546-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Immunoprecipitating (IP) PANX3 in MCF-7 whole cell lysate.
Western blot analysis of PANX3 using anti-PANX3 antibody (A11546-1);
Lane 1: MCF-7 whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-PANX3 antibody in MCF-7 whole cell lysate;
Lane 3: anti-PANX3 antibody (2μg) + MCF-7 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-PANX3 antigen affinity purified polyclonal antibody (A11546-1) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1196-200). A specific band was detected for PANX3 at approximately 45 kDa. The expected band size for PANX3 is at 45 kDa.