Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ATAD2 antibody [EPR12730] (ab176319)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR12730] to ATAD2
  • Suitable for: WB, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-ATAD2 antibody [EPR12730]
    See all ATAD2 primary antibodies

  • Description

    Rabbit monoclonal [EPR12730] to ATAD2

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human ATAD2 aa 50-150 (Cysteine residue). The exact sequence is proprietary.
    Database link: Q6PL18

  • Positive control

    • WB: HeLa, MCF-7, T47-D and Saos-2 cell lysates. ICC/IF: HeLa cells. 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-ATAD2 antibody [EPR12730] (ab176319)
    Western blot - Anti-ATAD2 antibody [EPR12730] (ab176319)

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: ATAD2 knockout HAP1 whole cell lysate (20 µg)

    Lanes 1 - 2: Merged signal (red and green). Green - ab176319 observed at 200 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab176319 was shown to specifically react with ATAD2 in wild type cells as signal was lost in ATAD2 knockout cells. Wild-type and ATAD2 knockout samples were subjected to SDS-PAGE. Ab176319 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

     

  • Flow Cytometry - Anti-ATAD2 antibody [EPR12730] (ab176319)
    Flow Cytometry - Anti-ATAD2 antibody [EPR12730] (ab176319)

    Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling ATAD2 (red) with ab176319 at a 1/60 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

  • Immunocytochemistry/ Immunofluorescence - Anti-ATAD2 antibody [EPR12730] (ab176319)
    Immunocytochemistry/ Immunofluorescence - Anti-ATAD2 antibody [EPR12730] (ab176319)

    Immunofluorescent staining of HeLa cells labeling ATAD2 using ab176319 at a 1/100 dilution.

  • Western blot - Anti-ATAD2 antibody [EPR12730] (ab176319)
    Western blot - Anti-ATAD2 antibody [EPR12730] (ab176319)
    All lanes : Anti-ATAD2 antibody [EPR12730] (ab176319) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ATAD2 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 159 kDa
    Observed band size: 200 kDa
    why is the actual band size different from the predicted?



    Lanes 1- 2: Merged signal (red and green). Green - ab176319 observed at 200 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

     ab176319 was shown to react with ATAD2 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab264957 (knockout cell lysate ab257359) lane below 200kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and ATAD2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab176319 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 in 1000 dilution and a 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-ATAD2 antibody [EPR12730] (ab176319)
    Western blot - Anti-ATAD2 antibody [EPR12730] (ab176319)
    All lanes : Anti-ATAD2 antibody [EPR12730] (ab176319) at 1/1000 dilution

    Lane 1 : HeLa lysate
    Lane 2 : MCF-7 lysate
    Lane 3 : T47-D lysate
    Lane 4 : Saos-2 lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 159 kDa

  • Anti-ATAD2 antibody [EPR12730] (ab176319)
    Anti-ATAD2 antibody [EPR12730] (ab176319)

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