Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-GRK2 antibody [EPR22465] (ab227825)

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Overview

  • 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 within Human GRK2 aa 100 to the C-terminus. The exact sequence is proprietary.
    Database link: P25098

  • 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.

  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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
WB 1/1000. Predicted molecular weight: 80 kDa.
IP 1/30.
  • Application notes
    Is unsuitable for ICC/IF or IHC-P.

    • Target

    • 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

    • Database links

      • Alternative names

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

      Images

      • Western blot - Anti-GRK2 antibody [EPR22465] (ab227825)
        Western blot - Anti-GRK2 antibody [EPR22465] (ab227825)
        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 kDa



        Blocking/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.

      • Western blot - Anti-GRK2 antibody [EPR22465] (ab227825)
        Western blot - Anti-GRK2 antibody [EPR22465] (ab227825)
        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 minutes


        Blocking/Dilution buffer: 5% NFDM/TBST.

      • Immunoprecipitation - Anti-GRK2 antibody [EPR22465] (ab227825)
        Immunoprecipitation - Anti-GRK2 antibody [EPR22465] (ab227825)

        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.

      • Immunoprecipitation - Anti-GRK2 antibody [EPR22465] (ab227825)
        Immunoprecipitation - Anti-GRK2 antibody [EPR22465] (ab227825)

        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.

      References

      ab227825 has not yet been referenced specifically in any publications.

      Publishing research using ab227825? Please let us know so that we can cite the reference in this datasheet.

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