Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-GRK2 antibody [EPR22465] (ab227825)

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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

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

    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
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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 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 kDa



        Lanes 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.

      • 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.

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

      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.

      Click here to view the general protocols

      References (0)

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

      ab227825 has not yet been referenced specifically in any publications.

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