Anti-PKC alpha/PRKCA Antibody Picoband®

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.

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.

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.

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.

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.