Invitrogen

GRK2 Monoclonal Antibody (3F8)

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated. View Details

1 Published Figure

1 Reference

View all (24) GRK2 antibodies

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Datasheet

Protocols

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GRK2 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of GRK2 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with GRK2 Monoclonal Antibody (3F8) (Product # MA5-15840) at 1:250 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

GRK2 Antibody in Immunohistochemistry (IHC) — Immunohistochemical analysis of paraffin-embedded endometrial cancer tissues (left) and cervical cancer tissues (right) using GRK2 monoclonal antibody (Product # MA5-15840) followed with DAB staining.

GRK2 Antibody in Western Blot (WB) — Knockdown of GRK2 was achieved by transfecting HEK-293 cells with GRK2 specific siRNAs (Silencer® select Product # s1129). Western blot analysis (Fig. a) was performed using whole cell extracts from the GRK2 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with GRK2 Monoclonal Antibody (Product # MA5-15840, 1:2000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to GRK2.

GRK2 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HEK-293 cells were transfected with GRK2 siRNA and reduction of signal was observed in Western Blot using GRK2 Monoclonal Antibody (Product # MA5-15840). {KD}

GRK2 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Fig 5 ADRBK1 overexpression promotes growth in human PDAC. A: Box-and-whisker plot showing ADRBK1 mRNA expression in primary human pancreatic tumor tissue samples, chronic pancreatitis and normal pancreas as analyzed by quantitative realtime reverse transcription PCR (qRT-PCR). Expression was normalized to ribosomal protein, large, P0 (RPLP0) mRNA levels. Data in Fig. represent median and 2nd and 3rd quartiles (boxes) as well as minimum and maximum values (whiskers). CP = Chronic Pancreatitis. ** p<0.01, *** p<0.001 (Student's t-test). B: Immunohistological analyses of ADRBK1 expression using human pancreatic tissue microarrays (TMAs) comprising 33 donor, 28 chronic pancreatitis and 65 tumor tissues. Micrographs are representative of typical staining results for donor samples (left panel), which were devoid of ADRBK1 staining, or ADRBK1-positive neoplastic tissues (right panel) (original magnification 100x). Note that in positive PDAC cases, neoplastic ducts as well as individual infiltrating immune cells (arrows) stained postitve for ADRBK1. Lower panel: Distribution of staining intensities across different tumor grades (G1: n = 23; G2: n = 19; G3: n = 23). C: TTK mRNA expression levels in a variety of different pancreatic cancer and control cell lines (qRT-PCR; expression was normalized to ribosomal protein, large, P0 (RPLP0) mRNA levels). D, E: MTT assays demonstrated increased numbers of viable S2-007 (C) and Panc-1 (D) cells 72h after transfection of fluorescence-ta

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1:500-1:2,000

Immunohistochemistry (IHC)

1:200-1:1,000

Immunohistochemistry (Paraffin) (IHC (P))

View 1 publication 1 publication

Immunocytochemistry (ICC/IF)

1:200-1:1,000

ELISA (ELISA)

1:10,000

Product Specifications

Species Reactivity

Human, Mouse, Non-human primate, Rat

Published species

Human

Host/Isotype

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

3F8

Immunogen

Purified recombinant fragment of human GRK2 expressed in E. Coli.

Conjugate

Unconjugated

Form

Liquid

Concentration

Conc. Not Determined

Storage buffer

ascites

Contains

0.03% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_11152830

Product Specific Information

MA5-15840 targets GRK2 in IF, IHC, and WB applications and shows reactivity with Human, mouse, Non-human primate, and Rat samples.

The MA5-15840 immunogen is purified recombinant fragment of human GRK2 expressed in E. Coli. .

MA5-15840 detects GRK2 which has a predicted molecular weight of approximately 80kDa.

Target Information

G protein-coupled receptor kinases (GRKs) are important regulators of G protein-coupled receptors(GPCRs). GRK2 (83 kDa), one of six members of this family that have been identified, is ubiquitously expressed in mammals. After binding to their ligand and interacting with heterotrimeric G proteins, GPCRs (e.g., a2-adrenergic receptor) are phosphorylated by GRKs. Internalization of the GPCRs, regulated by beta-arrestin-1, leads to activation of the Ras - Raf - ERK1&2 signaling pathway. GRK2 activity is tightly controlled by different mechanisms including phosphorylation by kinases such as PKC, Src and ERK1&2, as well as interaction with various proteins. ERK phosphorylates and thus inactivates GRK2 on serine 670 in a negative feedback mechanism.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: adrenergic beta receptor kinase 1; adrenergic receptor kinase, beta 1; adrenergic, beta, receptor kinase 1; Beta-adrenergic receptor kinase 1; beta-adrenergic receptor kinase 1 beta ARK1; beta-AR kinase-1; Beta-ARK-1; EC 2.7.1.126; G-protein coupled receptor kinase 2; G-protein-coupled receptor kinase 2; grk 2

Gene Aliases: Adrbk-1; ADRBK1; BARK; Bark-1; BARK1; beta ARK; BETA-ARK1; betaARK1; GRK-2; GRK2

UniProt ID: (Mouse) Q99MK8, (Rat) P26817, (Human) P25098

Entrez Gene ID: (Mouse) 110355, (Rat) 25238, (Human) 156

Molecular Function: non-receptor serine/threonine protein kinase