Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

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 - BSA and Azide free
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Human

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 & Species

    Application Species
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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

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

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

Properties

Images

  • Western blot - Anti-GRK2 antibody [EPR22465] - BSA and Azide free (ab245125)
    Western blot - Anti-GRK2 antibody [EPR22465] - BSA and Azide free (ab245125)
    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



    This data was developed using ab227825, the same antibody clone in a different buffer formulation.

    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.

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

    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).

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

    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).

  • Western blot - Anti-GRK2 antibody [EPR22465] - BSA and Azide free (ab245125)
    Western blot - Anti-GRK2 antibody [EPR22465] - BSA and Azide free (ab245125)
    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).

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

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