Recombinant Anti-GRK2 antibody [EPR22465] (ab227825)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22465] to GRK2
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-GRK2 antibody [EPR22465]
See all GRK2 primary antibodies -
Description
Rabbit monoclonal [EPR22465] to GRK2 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IPmore details
Unsuitable for: ICC/IF or IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, HEK-293 and HepG2 whole cell lysates; Wild-type HAP1 whole cell lysate; Human skeletal muscle lysate. IP: HeLa and HEK-293T whole cell lysates.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22465 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab227825 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000. Predicted molecular weight: 80 kDa.
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IP |
1/30.
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Notes |
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WB
1/1000. Predicted molecular weight: 80 kDa. |
IP
1/30. |
Target
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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. -
Tissue specificity
Expressed in peripheral blood leukocytes. -
Sequence similarities
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. GPRK subfamily.
Contains 1 AGC-kinase C-terminal domain.
Contains 1 PH domain.
Contains 1 protein kinase domain.
Contains 1 RGS domain. - Information by UniProt
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Database links
- Entrez Gene: 156 Human
- Omim: 109635 Human
- SwissProt: P25098 Human
- Unigene: 83636 Human
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Alternative names
- ADRBK1 antibody
- Adrenergic beta receptor kinase 1 antibody
- ARBK1_HUMAN antibody
see all
Images
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All lanes : Anti-GRK2 antibody [EPR22465] (ab227825) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : ADRBK1 knockout HEK293T cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 80 kDa
Observed band size: 80 kDaLanes 1-4: Merged signal (red and green). Green - ab227825 observed at 80 kDa. Red - loading control ab8245 observed at 36 kDa.
ab227825 Anti-GRK2 antibody [EPR22465] was shown to specifically react with GRK2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266352 (knockout cell lysate ab257345) was used. Wild-type and GRK2 knockout samples were subjected to SDS-PAGE. ab227825 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-GRK2 antibody [EPR22465] (ab227825) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : GRK2 knockout HAP1 whole cell lysate
Lane 3 : HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate
Lane 4 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 5 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 80 kDaBlocking/Dilution buffer: NFDM/TBST.
ab227825 was shown to specifically react with GRK2 in wild-type HAP1 cells as signal was lost in GRK2 knockout cells. Wild-type and GRK2 knockout samples were subjected to SDS-PAGE. ab227825 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.
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Anti-GRK2 antibody [EPR22465] (ab227825) at 1/1000 dilution + Human skeletal muscle lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Predicted band size: 80 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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GRK2 was immunoprecipitated from 0.35 mg HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab227825 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab227825 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: HeLa whole cell lysate 10 µg (Input).
Lane 2: ab227825 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab227825 in HeLa whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 seconds.GRK2 was found readily degradable by proteolytic process (PMID:9857063; PMID:12738776). The bands smaller than 80-kDa detected in the immune-precipitate may represent degraded GRK2.
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GRK2 was immunoprecipitated from 0.35 mg HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate with ab227825 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab227825 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: HEK-293T whole cell lysate 10 µg (Input).
Lane 2: ab227825 IP in HEK-293T whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab227825 in HEK-293T whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 seconds.GRK2 was found readily degradable by proteolytic process (PMID:9857063; PMID:12738776). The bands smaller than 80-kDa detected in the immune-precipitate may represent degraded GRK2.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab227825 has not yet been referenced specifically in any publications.