Product: GRK2 Antibody
Catalog: DF6538
Description: Rabbit polyclonal antibody to GRK2
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Dog, Chicken
Mol.Wt.: 80kDa; 80kD(Calculated).
Uniprot: P25098
RRID: AB_2838500

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 100ul $280 In stock
 200ul $350 In stock

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Dog(100%), Chicken(90%)
Clonality:
Polyclonal
Specificity:
GRK2 Antibody detects endogenous levels of total GRK2.
RRID:
AB_2838500
Cite Format: Affinity Biosciences Cat# DF6538, RRID:AB_2838500.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

ADRBK1; Adrenergic beta receptor kinase 1; ARBK1_HUMAN; BARK; BARK1; Beta adrenergic receptor kinase 1; Beta ARK 1; Beta ARK1; Beta-adrenergic receptor kinase 1; Beta-ARK-1; FLJ16718; G protein coupled receptor kinase 2; G-protein coupled receptor kinase 2; GRK2;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P25098 ARBK1_HUMAN:

Expressed in peripheral blood leukocytes.

Description:
G-protein-coupled receptor kinase 2 (GRK2), also known as beta-adrenergic receptor kinase 1 (beta-ARK1), is a member of the GRK family, which phosphorylates the activated form of G-protein-coupled receptors (GPCRs) and initiates the desensitization process of GPCR (1). GRK2 kinase activity and cellular localization are tightly regulated by interactions with activated receptors, G-beta and G-gamma subunits, adaptor proteins, phospholipids, caveolin and calmodulin, as well as by phosphorylation (1). PKC phosphorylation enhances GRK2 activity by promoting its membrane localization and by abolishing the inhibitory association of calmodulin (2,3). PKA phosphorylates GRK2 at Ser685, which facilitates the association of GRK2 with a beta-adrenergic receptor (4). Erk inhibits GRK2 activity via phosphorylation at Ser670 (5). Src phosphorylates GRK2 at multiple tyrosine residues (Tyr13, 86 and 92), which activates GRK2 activity and promotes GRK2 degradation (6,7).
Sequence:
MADLEAVLADVSYLMAMEKSKATPAARASKKILLPEPSIRSVMQKYLEDRGEVTFEKIFSQKLGYLLFRDFCLNHLEEARPLVEFYEEIKKYEKLETEEERVARSREIFDSYIMKELLACSHPFSKSATEHVQGHLGKKQVPPDLFQPYIEEICQNLRGDVFQKFIESDKFTRFCQWKNVELNIHLTMNDFSVHRIIGRGGFGEVYGCRKADTGKMYAMKCLDKKRIKMKQGETLALNERIMLSLVSTGDCPFIVCMSYAFHTPDKLSFILDLMNGGDLHYHLSQHGVFSEADMRFYAAEIILGLEHMHNRFVVYRDLKPANILLDEHGHVRISDLGLACDFSKKKPHASVGTHGYMAPEVLQKGVAYDSSADWFSLGCMLFKLLRGHSPFRQHKTKDKHEIDRMTLTMAVELPDSFSPELRSLLEGLLQRDVNRRLGCLGRGAQEVKESPFFRSLDWQMVFLQKYPPPLIPPRGEVNAADAFDIGSFDEEDTKGIKLLDSDQELYRNFPLTISERWQQEVAETVFDTINAETDRLEARKKAKNKQLGHEEDYALGKDCIMHGYMSKMGNPFLTQWQRRYFYLFPNRLEWRGEGEAPQSLLTMEEIQSVEETQIKERKCLLLKIRGGKQFILQCDSDPELVQWKKELRDAYREAQQLVQRVPKMKNKPRSPVVELSKVPLVQRGSANGL

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Chicken
90
Xenopus
0
Zebrafish
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P25098 As Substrate

Site PTM Type Enzyme
Y13 Phosphorylation P00533 (EGFR) , P12931 (SRC)
S29 Phosphorylation P17252 (PRKCA) , P05129 (PRKCG) , Q05655 (PRKCD)
K31 Ubiquitination
K45 Ubiquitination
R50 Methylation
T54 Phosphorylation
K57 Ubiquitination
K62 Ubiquitination
Y86 Phosphorylation P00533 (EGFR) , P12931 (SRC)
Y92 Phosphorylation P12931 (SRC) , P00533 (EGFR)
K94 Ubiquitination
K115 Ubiquitination
K126 Ubiquitination
K138 Ubiquitination
K139 Ubiquitination
K164 Ubiquitination
K170 Ubiquitination
Y206 Phosphorylation
T213 Phosphorylation
K215 Ubiquitination
K220 Ubiquitination
K230 Ubiquitination
S247 Phosphorylation
T248 Phosphorylation
T263 Phosphorylation
K319 Ubiquitination
K344 Ubiquitination
K345 Ubiquitination
K346 Ubiquitination
S350 Phosphorylation
T353 Phosphorylation
Y356 Phosphorylation
S416 Phosphorylation
S423 Phosphorylation
K448 Ubiquitination
S487 Phosphorylation
K494 Ubiquitination
K497 Ubiquitination
S501 Phosphorylation
Y506 Phosphorylation
K545 Ubiquitination
K557 Ubiquitination
K628 Ubiquitination
K644 Acetylation
K644 Ubiquitination
S670 Phosphorylation P27361 (MAPK3) , P24941 (CDK2)
S676 Phosphorylation
K677 Ubiquitination
R683 Methylation
S685 Phosphorylation P17612 (PRKACA)

PTMs - P25098 As Enzyme

Substrate Site Source
P05387 (RPLP2) S102 Uniprot
P05387 (RPLP2) S105 Uniprot
P07550 (ADRB2) S355 Uniprot
P07550 (ADRB2) S356 Uniprot
P07550 (ADRB2) T384 Uniprot
P07550 (ADRB2) S396 Uniprot
P07550 (ADRB2) S401 Uniprot
P07550 (ADRB2) S407 Uniprot
P07550 (ADRB2) S411 Uniprot
P08069 (IGF1R) S1278 Uniprot
P08172 (CHRM2) S232 Uniprot
P08172 (CHRM2) S234 Uniprot
P08172 (CHRM2) S250 Uniprot
P08172 (CHRM2) S251 Uniprot
P08172 (CHRM2) T271 Uniprot
P08172 (CHRM2) S282 Uniprot
P08172 (CHRM2) S283 Uniprot
P08172 (CHRM2) S286 Uniprot
P08172 (CHRM2) T287 Uniprot
P08172 (CHRM2) S288 Uniprot
P08172 (CHRM2) S290 Uniprot
P08172 (CHRM2) S294 Uniprot
P08172 (CHRM2) T302 Uniprot
P08172 (CHRM2) T307 Uniprot
P08172 (CHRM2) S309 Uniprot
P08172 (CHRM2) T310 Uniprot
P08172 (CHRM2) S311 Uniprot
P09497 (CLTB) S205 Uniprot
P15311 (EZR) T567 Uniprot
P18545 (PDE6G) T62 Uniprot
P21731-2 (TBXA2R) S239 Uniprot
P21731-2 (TBXA2R) S357 Uniprot
P26038 (MSN) T558 Uniprot
P30411 (BDKRB2) S366 Uniprot
P30411 (BDKRB2) S373 Uniprot
P30411 (BDKRB2) S375 Uniprot
P32245 (MC4R) T312 Uniprot
P32245 (MC4R) S329 Uniprot
P32245 (MC4R) S330 Uniprot
P35241 (RDX) T564 Uniprot
P35367 (HRH1) S398 Uniprot
P35372 (OPRM1) T356 Uniprot
P35372 (OPRM1) S357 Uniprot
P35372 (OPRM1) S358 Uniprot
P35372 (OPRM1) S377 Uniprot
P37840 (SNCA) S129 Uniprot
P41143 (OPRD1) T358 Uniprot
P41143 (OPRD1) S363 Uniprot
Q14940 (SLC9A5) S702 Uniprot
Q14940 (SLC9A5) S709 Uniprot
Q14940 (SLC9A5) S711 Uniprot
Q14940 (SLC9A5) S712 Uniprot
Q14940 (SLC9A5) T714 Uniprot
Q15796 (SMAD2) T197 Uniprot
Q16539 (MAPK14) T123 Uniprot
Q96A54 (ADIPOR1) S7 Uniprot
Q96A54 (ADIPOR1) T24 Uniprot
Q96A54 (ADIPOR1) T53 Uniprot
Q9Y2W7 (KCNIP3) S95 Uniprot

Research Backgrounds

Function:

Specifically phosphorylates the agonist-occupied form of the beta-adrenergic and closely related receptors, probably inducing a desensitization of them. Key regulator of LPAR1 signaling. Competes with RALA for binding to LPAR1 thus affecting the signaling properties of the receptor. Desensitizes LPAR1 and LPAR2 in a phosphorylation-independent manner. Positively regulates ciliary smoothened (SMO)-dependent Hedgehog (Hh) signaling pathway by facilitating the trafficking of SMO into the cilium and the stimulation of SMO activity (By similarity).

Subcellular Location:

Cytoplasm. Cell membrane.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed in peripheral blood leukocytes.

Subunit Structure:

Interacts with GIT1 (By similarity). Interacts with, and phosphorylates chemokine-stimulated CCR5. Interacts with ARRB1. Interacts with LPAR1 and LPAR2. Interacts with RALA in response to LPAR1 activation. ADRBK1 and RALA mutually inhibit each other's binding to LPAR1. Interacts with ADRB2.

Family&Domains:

The PH domain binds anionic phospholipids and helps recruiting ADRBK1 from the cytoplasm to plasma membrane close to activated receptors. It mediates binding to G protein beta and gamma subunits, competing with G-alpha subunits and other G-betagamma effectors.

Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. GPRK subfamily.

References

1). Angiotensin II Type 2 Receptor Modulates Synovial Macrophage Polarization by Inhibiting GRK2 Membrane Translocation in a Rat Model of Collagen-Induced Arthritis. JOURNAL OF IMMUNOLOGY (PubMed: 33148713) [IF=4.4]

Application: WB    Species: rat    Sample: Bone marrow–derived macrophages

FIGURE 9. AT2R activation inhibits GRK2 membrane translocation and promotes the coexpression with p-ERK1/2. BMDMs were pretreated with CGP42112 or PD123319 before the addition of LPS (1 mg/ml, 4 h). (A) The membrane protein levels of GRK2 were determined by flow cytometry (n = 5). (B) The total, membrane, and cytoplasm protein levels of GRK2 were determined by Western blot analyses (n = 4). (C) Confocal imaging of GRK2 translocation in BMDMs, Original magnification 3400. (D) LPS-stimulated BMDMs with CGP42112 or PD123319 were immunoprecipitated using an anti-GRK2 Ab and subjected to Western blotting to visualize ERK1/2 to evaluate its association with the GRK2 in cytoplasm. Coimmunoprecipitation (IP): anti-GRK2; immunoblot (IB): p-ERK1/2 and GRK2. (n = 3). **p , 0.01 versus control group; #p , 0.05, ##p , 0.01 versus LPS group; $p , 0.05, $$p , 0.01 versus LPS plus CGP group.

2). The ROS/GRK2/HIF-1α/NLRP3 Pathway Mediates Pyroptosis of Fibroblast-Like Synoviocytes and the Regulation of Monomer Derivatives of Paeoniflorin. Oxidative Medicine and Cellular Longevity (PubMed: 35132350)

Application: WB    Species: Human    Sample:

Figure 5 G protein-coupled receptor kinase 2 (GRK2) promotes the expression of hypoxia-inducible factor (HIF)-1α. (a, b) shGRK2 was transfected into fibroblast-like synoviocytes (FLS) and exposed to hypoxia for 2 h. Immunoblot analysis and quantification of GRK2 and HIF-1α. (c, d) GRK2 overexpression plasmid was transfected into FLS. Immunoblot analysis and quantification of GRK2, HIF-1α, nucleotide-binding oligomerization segment-like receptor family 3 (NLRP3), and N-terminal domain of human gasdermin D (GSDMD-N). The values are shown as the mean ± standard deviation (n = 3). ∗p < 0.05, ∗∗p < 0.01 compared with the control group and #p < 0.05 compared with the hypoxia group or the NC group.

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