Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ADAR1 antibody [EPR7033] (ab126745)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR7033] to ADAR1
  • Suitable for: WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-ADAR1 antibody [EPR7033]
    See all ADAR1 primary antibodies

  • Description

    Rabbit monoclonal [EPR7033] to ADAR1

  • Host species

    Rabbit

  • Specificity

    The immunogen is designed to detect the p150 isoform and not the p110.

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human ADAR1 aa 200-300. The exact sequence is proprietary. The immunogen used to raise this antibody is designed to detect isoform 1 (p150) and isoforms 2-4. It does not detect Isoform 5 (p110).
    Database link: P55265

  • Positive control

    • WB: HEK293T, HeLa, Ramos and SH-SY5Y cell lysates. IHC-P: Human brain tissue Flow Cyt: HeLa cells

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Images

  • Western blot - Anti-ADAR1 antibody [EPR7033] (ab126745)
    Western blot - Anti-ADAR1 antibody [EPR7033] (ab126745)
    All lanes : Anti-ADAR1 antibody [EPR7033] (ab126745) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : ADAR knockout HEK293T cell lysate
    Lane 3 : 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: 136 kDa
    Observed band size: 130 kDa
    why is the actual band size different from the predicted?



    Lanes 1-3: Merged signal (red and green). Green - ab126745 observed at 130 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab126745 Anti-ADAR1 antibody [EPR7033] was shown to specifically react with ADAR1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266846 (knockout cell lysate ab257131) was used. Wild-type and ADAR1 knockout samples were subjected to SDS-PAGE. ab126745 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADAR1 antibody [EPR7033] (ab126745)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADAR1 antibody [EPR7033] (ab126745)

    ab126745, at 1/50 dilution, staining ADAR1 in paraffin-embedded Human brain tissue by Immunohistochemistry.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-ADAR1 antibody [EPR7033] (ab126745)
    Flow Cytometry - Anti-ADAR1 antibody [EPR7033] (ab126745)
    Flow cytometric analysis of permeabilized Ramos cells, staining ADAR1 (red) with ab126745.

    1x106 cells were collected and washed with blocking buffer. Cells were fixed with 2% paraformaldehyde, permeabilized with 1X FACS permeabilizing solution and blocked with blocking buffer for 30 minutes at room temperature. Cells were incubated with primary antibody (1/10) for 30 minutes at room temperature before a fluorescently-conjugated secondary antibody or 30 min at room temperature. A rabbit IgG was used as a negative control (green).
  • Western blot - Anti-ADAR1 antibody [EPR7033] (ab126745)
    Western blot - Anti-ADAR1 antibody [EPR7033] (ab126745)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: ADAR1 knockout HAP1 cell lysate (20 µg)
    Lane 3: HepG2 cell lysate (20 µg)
    Lane 4: HeLa cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab126745 observed at 150 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab126745 was shown to recognize ADAR1 when ADAR1 knockout samples were used, along with additional cross-reactive bands. Wild-type and ADAR1 knockout samples were subjected to SDS-PAGE. ab126745 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging.

  • Western blot - Anti-ADAR1 antibody [EPR7033] (ab126745)
    Western blot - Anti-ADAR1 antibody [EPR7033] (ab126745)
    All lanes : Anti-ADAR1 antibody [EPR7033] (ab126745) at 1/1000 dilution

    Lane 1 : HeLa (treated with IFN-alpha) cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : Ramos cell lysate
    Lane 4 : SH-SY5Y cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 136 kDa
    Observed band size: 150 kDa why is the actual band size different from the predicted?

  • Anti-ADAR1 antibody [EPR7033] (ab126745)
    Anti-ADAR1 antibody [EPR7033] (ab126745)

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