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Anti-p53 antibody [PAb 1801] (ab28)

Key features and details

  • Mouse monoclonal [PAb 1801] to p53
  • Suitable for: WB, ELISA
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-p53 antibody [PAb 1801]
    See all p53 primary antibodies

  • Description

    Mouse monoclonal [PAb 1801] to p53

  • Host species

    Mouse

  • Tested Applications & Species

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

  • Immunogen

    Fusion protein corresponding to Human p53 (N terminal).
    Database link: P04637

  • Epitope

    aa 46-55 of human p53

  • Positive control

    • WB: A431 cell lysate, MDA-MB-231 cell lysate.

  • General notes

    This monoclonal p53 antibody has been knockout validated in Western blot. The expected band for p53 was observed in HCT116 cells treated with irinotecan and the band was not seen in TP53 knockout HCT116 cell lysate.

    For Western blot, we recommend using 3% milk as the blocking agent.

    We recommend using ab1101 or ab154036, alternative mouse monoclonal antibodies, to detect human p53 by IHC, ICC/IF or flow cytometry since they perform better in these applications.

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Images

  • Western blot - Anti-p53 antibody [PAb 1801] (ab28)
    Western blot - Anti-p53 antibody [PAb 1801] (ab28)
    All lanes : Anti-p53 antibody [PAb 1801] (ab28) at 5 µg/ml

    Lane 1 : Wild-type HCT116 cell lysate at 30 µg
    Lane 2 : Wild-type HCT116 + irinotecan (10 µM, 24 hours) cell lysate at 30 µg
    Lane 3 : p53 knockout HCT116 cell lysate at 30 µg
    Lane 4 : p53 knockout HCT116 + irinotecan (10 µM, 24 hours) cell lysate at 30 µg
    Lane 5 : A431 cell lysate (positive control) at 20 µg
    Lane 6 : Saos-2 cell lysate (negative control) at 20 µg
    Lane 7 : MEF cell lysate at 20 µg
    Lane 8 : Wild-type HAP1 cell lysate at 20 µg
    Lane 9 : p53 knockout HAP1 cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 53 kDa



    Lanes 1-9: Merged (red and green) signal.


    Ab28 was shown to specifically react with p53 in wild-type HAP1 and HCT116 cells treated with irinotecan. No band was observed in p53 knockout HAP1 or HCT116 cells. Wild-type and p53 knockout samples, positive and negative controls were subjected to SDS-PAGE. Ab28 and ab181602(loading control to GAPDH) were diluted 5 μg/mL and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging.

  • Western blot - Anti-p53 antibody [PAb 1801] (ab28)
    Western blot - Anti-p53 antibody [PAb 1801] (ab28)
    Anti-p53 antibody [PAb 1801] (ab28) at 5 µg/ml + MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 20 µg

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 53 kDa
    Additional bands at: 53 kDa, 60 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes


    This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab28 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.

     

    We recommend using 3% milk as the blocking agent for Western blot.

  • ELISA - Anti-p53 antibody [PAb 1801] (ab28)
    ELISA - Anti-p53 antibody [PAb 1801] (ab28)
    Analysis of recombinant p53 protein (ab43615) in MCF7 cells treated with vehicle (VEH)as well as MCF7 cells treated with camptothecin (CAM) and no material (NO) by sandwich ELISA with the use of anti-53 antibody (ab28) as capture and anti-p53 antibody (ab32389) as detector.
  • Western blot - Anti-p53 antibody [PAb 1801] (ab28)
    Western blot - Anti-p53 antibody [PAb 1801] (ab28)

    All Lanes: Anti-p53 antibody [PAb 1801] (ab28) at 5 µg/ml

    Lane 1 and 10: Wild-type HCT116 cell lysate

    Lane 2 and 11: Wild-type HCT116 + irinotecan (10 µM, 24 hours) cell lysate

    Lane 3 and 12: p53 knockout HCT116 cell lysate

    Lane 4 and 13: p53 knockout HCT116 + irinotecan (10 µM, 24 hours) cell lysate

    Lane 5 and 14: A431 cell lysate (positive control)

    Lane 6 and 15: Saos-2 cell lysate (negative control)

    Lane 7 and 16: MEF cell lysate

    Lane 8 and 17: Wild-type HAP1 cell lysate

    Lane 9 and 18: p53 knockout HAP1 cell lysate

    Lanes 1-9: 1% BSA blocking buffer

    Lanes 10-18: 3% Milk blocking buffer

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) at 1/5000 dilution

    We recommend using 3% milk as the blocking agent for western blot.

  • Western blot - Anti-p53 antibody [PAb 1801] (ab28)
    Western blot - Anti-p53 antibody [PAb 1801] (ab28)

    The image shows a Western blot for ab28, testing the threshold detection of the p53 antibody. The lanes are as follows:

    Lane 1 - 20ug of A549 lysate, lane 2 - 10ug of A549 lysate, lane 3 - 5ug of A549 lysate, lane 4 - 2.5ug of A549 lysate, lane 5 - 1.5ug of A549 lysate.

    This picture was submitted as part of the review by Craig Carson.

     

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