Analysis Note

ControlHuman breast carcinoma tissue, mouse heart tissue, SK OV 3 cell lysates and NIH 3T3 cell lysates.
Included Positive Antigen Control: Catalog # 12-305, 3T3/A31 cell lysate. Add 2.5 µL of 2-mercaptoethanol/100 µL of lysate and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced lysate per lane for minigels.

Application

Research CategorySignaling

Immunoprecipitation:
A previous lot of this antibody was used to immunoprecipitate GRK2/3 from transfected cells, as shown by an independent laboratory.

Research Sub CategoryGPCR, cAMP/cGMP & Calcium Signaling

Detect GRK 2/3 (βARK 1/2) using this Anti-GRK 2/3 (βARK 1/2) Antibody, clone C5/1.1 validated for use in IP & WB.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

The beta-adrenergic receptor kinase specifically phosphorylates the agonist-occupied form of the beta-adrenergic and related G protein-coupled receptors. Overall, the ADRBK2 enzyme, also known as GRK3, has 85% amino acid similarity with ADRBK1, with the protein kinase catalytic domain having 95% similarity. The ADRBK2 mRNA is approximately 8 kilobases with a distribution similar to that of ADRBK1. These data suggest the existence of a family of receptor kinases which may serve broadly to regulate receptor function.
GRK2 phosphorylates the beta2 adrenergic receptor. It is a ubiquitous cytosolic enzyme that specifically phosphorylates the activated form of the beta-adrenergic and related G-protein-coupled receptors.

Immunogen

Partial GST fusion protein corresponding to the GRK3 (βARK2) carboxyl terminus (a.a. 467-688). This immunogen shares homology with GRK2 (βARK1). The minimal epitope recognized by the antibody is a.a. 483-485. Clone C5/1.1.

Epitope: a.a. 483-485

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Purified mouse monoclonal IgG2aκ in buffer containing 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl containing 0.05% sodium azide. Frozen solution.

Protein G Purified

Format: Purified

Quality

Routinely evaluated by immunoblot on RIPA lysates from mouse 3T3/A31 fibroblasts, bovine brain cytosol preparation, rat L6 myoblasts, human A431 carcinoma or human HeLa cell lysates.

Western Blot Analysis:
0.1-1 µg/mL of this lot detected GRK2/3 (βARK 1/2) in RIPA lysates from mouse 3T3/A31 fibroblasts. Previous lots of this antibody detected GRK2/3 in bovine brain cytosol preparation, rat L6 myoblasts, human A431 carcinoma and human HeLa cell RIPA lysates.

Specificity

This antibody recognizes GRK2/3, Mr 80 kDa. Additional bands at 85 and 50 kDa are also detected when using higher concentrations of the antibody.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.

Handling Recommendations:
Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

80 kDa