Anti-IKK gamma/IKBKG Antibody Picoband™

Figure 1. Western blot analysis of IKK gamma using anti-IKK gamma antibody (A00874).
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: rat cardiac muscle tissue lysates,
lane 2: HEPG2 whole cell 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 rabbit anti-IKK gamma antigen affinity purified polyclonal antibody (Catalog # A00874) 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 1:10000 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 IKK gamma at approximately 48KD. The expected band size for IKK gamma is at 48KD.

Figure 2. Flow Cytometry analysis of U87 cells using anti-IKK gamma antibody (A00874).
Overlay histogram showing U87 cells stained with A00874 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-IKK gamma Antibody (A00874,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 3. Flow Cytometry analysis of PC-3 cells using anti-IKK gamma antibody (A00874).
Overlay histogram showing PC-3 cells stained with A00874 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-IKK gamma Antibody (A00874,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 4. IF analysis of IKK gamma using anti-IKK gamma antibody (A00874).
IKK gamma was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-IKK gamma Antibody (A00874) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.