Anti-PKC alpha/PRKCA Antibody Picoband®

Figure 1. Western blot analysis of PKC alpha using anti-PKC alpha antibody (PB9318).
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 U87 whole cell lysates,
Lane 2: human HEL whole cell lysates,
Lane 3: human A549 whole cell lysates,
Lane 4: human U20S whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: mouse brain tissue 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-PKC alpha antigen affinity purified polyclonal antibody (Catalog # PB9318) 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 PKC alpha at approximately 77 kDa. The expected band size for PKC alpha is at 80 kDa.

Figure 2. IHC analysis of PKC alpha using anti-PKC alpha antibody (PB9318).
PKC alpha was detected in paraffin-embedded section of mouse intestine 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-PKC alpha Antibody (PB9318) 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 PKC alpha using anti-PKC alpha antibody (PB9318).
PKC alpha was detected in paraffin-embedded section of rat intestine 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-PKC alpha Antibody (PB9318) 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 PKC alpha using anti-PKC alpha antibody (PB9318).
PKC alpha was detected in paraffin-embedded section of human mammary 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-PKC alpha Antibody (PB9318) 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. Western blot analysis of PKC alpha/PRKCA using anti-PKC alpha/PRKCA Antibody (PB9318).
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.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 2.5% BSA on 1X-TBST 1.5 hour at RT. The membrane was incubated with rabbit anti-PKC alpha/PRKCA Antibody (PB9318) at 1:1000 (in 0.5%l BSA containing 1X-TBST) and overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a donkey anti-rabbit Ig-Cy5 secondary antibody at a dilution of 1:1000 (in 0.5%l BSA containing 1X-TBST) at 25°C for 2 h. The signal is developed using an CyDye. A specific band was detected for PKC alpha/PRKCA at approximately 76 kDa. The expected band size for PKC alpha/PRKCA is at 76 kDa.