Anti-GRK5 Antibody Picoband®

Western blot analysis of GRK5 using anti-GRK5 antibody (PB9708).
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 HepG2 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human A549 whole cell lysates,
Lane 4: rat heart tissue lysates,
Lane 5: mouse heart 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-GRK5 antigen affinity purified polyclonal antibody (Catalog # PB9708) 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 GRK5 at approximately 68 kDa. The expected band size for GRK5 is at 68 kDa.

IHC analysis of GRK5 using anti-GRK5 antibody (PB9708).
GRK5 was detected in a paraffin-embedded section of rat cardiac muscle tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-GRK5 Antibody (PB9708) 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 GRK5 using anti-GRK5 antibody (PB9708).
GRK5 was detected in a paraffin-embedded section of mouse lung tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-GRK5 Antibody (PB9708) 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 GRK5 using anti-GRK5 antibody (PB9708).
GRK5 was detected in a paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-GRK5 Antibody (PB9708) 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 GRK5 using anti-GRK5 antibody (PB9708).
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 5%(w/v) milk in TBST for 1.5 hour at RT. The membrane was incubated with rabbit anti-GRK5 antibody (PB9708) at 1 ug/ml overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with an anti-rabbit IgG(cell signaling,#7074) at 1:1000 for 2 hour at RT. The signal is developed using an Kodak Image Station 4000R Pro;Thermo Scientific West Femto Maximum Sensitivity Substrate(#34096). A specific band was detected for GRK5 at approximately 68 kDa. The expected band size for GRK5 is at 68 kDa.