Application

Western Blotting Analysis: A representative lot detected EGF-induced interaction between co-expressed bovine GRK2-GFP and FLAG-tagged human EGFR in HEK293 transfectants by Western blotting after anti-FLAG IP. Similar induction was observed when expressing catalytically inactive GRK2 K220R mutant or GRK2 lacking N-terminal 185 residues (Gao, J., et al. (2005). FEBS Lett. 579(1):122-126).
Western Blotting Analysis: A representative lot detected delta opioid receptor agonists- (DPDPE & DADLE) induced interaction between co-expressed bovine GRK2 and HA-tagged full-length murine delta opioid receptor (DOR), but not HA-mDOR lacking C-terminal end 31 residues, in HEK293 transfectants by Western blotting after anti-HA IP. Naloxone co-treatment abolished DPDPE effect (Li, J. et al. (2003). J. Biol. Chem. 278(32):30219-30226).
Western Blotting Analysis: A representative lot detected the expression of virally transfected full-length bovine GRK2 and C-terminal fragment (GRK2ct) in rabbit smooth muscle cells (Peppel, K., et al. (2000). Circulation. 102(7):793-799).
ELISA Analysis: Clone E23/8 was biotinylated and employed as the detection antibody for the detection of recombinant bovine GRK2 as well as endogenous GRK2 in human CCR5-expressing rat basophilic leukemia RBL-2H3 cells (RBL-CCR5), COS cells, and human leukocytes (lymphocytes, monocytes, granulocytes) by sandwich ELISA (Oppermann, M., et al. (1999). J. Biol. Chem. 274:8875 -8885).

This Anti-GRK2 Antibody, clone E23/8 is validated for use in Western Blotting, ELISA for the detection of GRK2.

General description

Beta-adrenergic receptor kinase 1 (UniProt P21146; EC 2.7.11.15; also known as Beta-ARK-1, G-protein-coupled receptor kinase 2) is encoded by the ADRBK1 (also known as GRK2) gene (Gene ID 282682) in bovine species. GRK2 belongs to the family of G-protein-coupled receptor (GPCR) kinases (GRKs) that are subdivided based on their structural and functional similarities into rhodopsin kinase (GRK1 and GRK7), the β-adrenergic receptor (βAR) kinase (GRK2 and GRK3), and the GRK4 (GRK4, GRK5, and GRK6) types. GRKs preferentially catalyze the phosphorylation of activated (agonist-occupied) rather than the inactive or antagonist-occupied GPCRs. The βAR kinase GRKs (GRK2 and GRK3) become associated with cell membranes by interacting with the βγ subunits of heterotrimeric G proteins (Gβγ) in a GPCR agonist-dependent manner, allowing subsequent phosphorylation of their target receptors.

Immunogen

BSA-conjugated linear peptide corresponding to the C-terminal sequence of bovine GRK2.

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Format: Purified

Quality

Evaluated by Western Blotting in lysate from bovine GRK2-expressing HEK293 cells.

Western Blotting Analysis: A 1:250 dilution of this antibody detected the exogenously expressed bovine GRK2 in lysate from transfected HEK293 cells.

Specificity

Clone E23/8 targets a GRK2 C-terminal epitope not present in GRK3. Clone E23/8 does not inhibit GRK2 activity or affect CCR5 phosphorylation in permeabilized RBL-CCR5 cells (Oppermann, M., et al. (1999). J. Biol. Chem. 274:8875 -8885).

Target description

~79 kDa observed. 79.65 kDa (bovine), 79.57 kDa (human), 79.64 kDa (mouse), 79.79 kDa (rat) calculated. Uncharacterized band(s) may appear in some lysates.