Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-GRK2 antibody [EPR22465] - BSA and Azide free (ab245125)

Overview

  • Product name

    Anti-GRK2 antibody [EPR22465] - BSA and Azide free
    See all GRK2 primary antibodies
  • Description

    Rabbit monoclonal [EPR22465] to GRK2 - BSA and Azide free
  • 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. IP: HeLa and HEK-293T whole cell lysates.
  • General notes

    Ab245125 is the carrier-free version of ab227825. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab245125 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 ab245125 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 Use at an assay dependent concentration. Predicted molecular weight: 80 kDa.
IP Use at an assay dependent concentration.
  • 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

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 secondary antibody (HRP) (ab131366), was used as secondary antibody 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227825).

    • All lanes : Anti-GRK2 antibody [EPR22465] - BSA and Azide free (ab245125) 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227825).

    References

    ab245125 has not yet been referenced specifically in any publications.

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