Anti-alpha 1d Adrenergic Receptor/ADRA1A Antibody Picoband®

Figure 1. Western blot analysis of ADRA1A using anti-ADRA1A antibody (PB9752).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: Rat Cardiac Muscle Tissue Lysate at 50ug,
Lane 2: Rat Brain Tissue Lysate at 50ug,
Lane 3: Rat Liver Tissue Lysate at 50ug,
Lane 4: Mouse Liver Tissue Lysate at 50ug,
Lane 5: Mouse Lung Tissue Lysate at 50ug,
Lane 6: 22RV1 Whole Cell Lysate at 40ug,
Lane 7: SMMC Whole Cell Lysate at 40ug.
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-ADRA1A antigen affinity purified polyclonal antibody (Catalog # PB9752) 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 ADRA1A at approximately 51 kDa. The expected band size for ADRA1A is at 51 kDa.

Figure 2. IHC analysis of ADRA1A using anti-ADRA1A antibody (PB9752).
ADRA1A was detected in paraffin-embedded section of human liver cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ADRA1A Antibody (PB9752) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 3. IHC analysis of ADRA1A using anti-ADRA1A antibody (PB9752).
ADRA1A was detected in paraffin-embedded section of rat brain tissue tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ADRA1A Antibody (PB9752) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 4. IHC analysis of ADRA1A using anti-ADRA1A antibody (PB9752).
ADRA1A was detected in paraffin-embedded section of rat brain tissue tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ADRA1A Antibody (PB9752) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 5. Flow Cytometry analysis of A431 cells using anti-ADRA1A antibody (PB9752).
Overlay histogram showing A431 cells stained with PB9752 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ADRA1A Antibody (PB9752,1μg/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.