GRK2 Antibody

• Catalog Number: orb669177
• Category: Antibodies
• Description: GRK2 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, WB
• Reactivity: Human, Monkey, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: A synthetic peptide corresponding to a sequence of human GRK2 (KKELRDAYREAQQLVQRV).
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.25-0.5μg/ml, Human, Mouse, Rat, Monkey Immunocytochemistry/Immunofluorescence, 5μg/ml, Human Flow Cytometry, 1-3μg/1x106 cells, Human, Mouse, Rat
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 80 kDa
• UniProt ID: P25098
• Storage: Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
• Note: For research use only
• Application notes: Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of GRK2 using anti-GRK2 antibody (orb669177). 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: human THP-1 whole cell lysates, Lane 2: monkey COS-7 whole cell lysates, Lane 3: human Raji whole cell lysates, Lane 4: rat lung tissue lysates, Lane 5: mouse NTH/3T3 whole cell lysate, Lane 6: mouse Raw264.7 whole cell lysate. 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-GRK2 antigen affinity purified polyclonal antibody (Catalog # orb669177) 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 # orb90503) with Tanon 5200 system. A specific band was detected for GRK2 at approximately 80KD. The expected band size for GRK2 is at 80KD.

IF analysis of GRK2 using anti-GRK2 antibody (orb669177). GRK2 was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-GRK2 Antibody (orb669177) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG 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.

Flow Cytometry analysis of HEPA1-6 cells using anti-GRK2 antibody (orb669177). Overlay histogram showing HEPA1-6 cells stained with orb669177 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-GRK2 Antibody (orb669177, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Flow Cytometry analysis of RH35 cells using anti-GRK2 antibody (orb669177). Overlay histogram showing RH35 cells stained with orb669177 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-GRK2 Antibody (orb669177, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Flow Cytometry analysis of U937 cells using anti-GRK2 antibody (orb669177). Overlay histogram showing U937 cells stained with orb669177 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-GRK2 Antibody (orb669177, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.