Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-p53 antibody [EPR17343] (ab179477)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17343] to p53
  • Suitable for: WB, ICC/IF
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-p53 antibody [EPR17343]
    See all p53 primary antibodies

  • Description

    Rabbit monoclonal [EPR17343] to p53

  • Host species

    Rabbit

  • Specificity

    Based on the results of the knockout validation testing this antibody may not be suitable for IHC-P, Flow Cyt, IP or ChIP. Please contact our Scientific Support team for additional information.

  • Tested Applications & Species

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

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A431, HEK-293, and T-47D whole cell lysates. ICC/IF: Wild-type HAP1 cells.

  • General notes

    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-p53 antibody [EPR17343] (ab179477)
    Western blot - Anti-p53 antibody [EPR17343] (ab179477)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)

    Lane 2: p53 knockout HAP1 cell lysate (20 µg)

    Lane 3: A431 cell lysate (20 µg)

    Lane 4: Saos-2 cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab179477 observed at 53 kDa. Red - loading control, ab8226, observed at 42 kDa.

    ab179477 was shown to specifically react with p53 in wild type HAP1 cells along with additional cross reactive bands. No band was observed with p53 knockout samples were used.

    Wild-type and p53 knockout samples were subjected to SDS-PAGE. ab179477 and ab8226 (loading control to beta Actin) were diluted to 1/2000 and 1/1000 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [EPR17343] (ab179477)
    Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [EPR17343] (ab179477)

    ab179477 staining p53 in wild-type HAP1 cells (top panel) and p53 knockout HAP1 cells (bottom panel). The cells were fixed with methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab179477 at 1/500 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

  • Western blot - Anti-p53 antibody [EPR17343] (ab179477)
    Western blot - Anti-p53 antibody [EPR17343] (ab179477)
    All lanes : Anti-p53 antibody [EPR17343] (ab179477) at 1/2000 dilution

    Lane 1 : Untreated HCT 116 (Human colorectal carcinoma) whole cell lysate
    Lane 2 : HCT 116 (Human colorectal carcinoma) whole cell lysate with 0.5µM Doxorubicin for 24hours whole cell lysate

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 44 kDa
    Observed band size: 53 kDa
    why is the actual band size different from the predicted?

  • Western blot - Anti-p53 antibody [EPR17343] (ab179477)
    Western blot - Anti-p53 antibody [EPR17343] (ab179477)
    Anti-p53 antibody [EPR17343] (ab179477) at 1/20000 dilution + T-47D (Human ductal breast epithelial carcinoma cell line) whole cell lysates at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 44 kDa
    Observed band size: 44,53 kDa why is the actual band size different from the predicted?



    Blocking/dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-p53 antibody [EPR17343] (ab179477)
    Western blot - Anti-p53 antibody [EPR17343] (ab179477)
    All lanes : Anti-p53 antibody [EPR17343] (ab179477) at 1/2000 dilution

    Lane 1 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysates
    Lane 2 : A431 (Human epidermoid carcinoma) whole cell lysates
    Lane 3 : Saos-2 (Human osteosarcoma cell line) whole cell lysates
    Lane 4 : HL-60 (Human promyelocytic leukemia cells) whole cell lysates
    Lane 5 : PC-3 (Human prostate adenocarcinoma cell line) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 44 kDa
    Observed band size: 53, 44 kDa why is the actual band size different from the predicted?



    Blocking and Diluting buffer and concentration: 5% NFDM/TBST.

    Saos-2, PC-3 and HL-60 cells are p53 null cell lines.

  • Anti-p53 antibody [EPR17343] (ab179477)
    Anti-p53 antibody [EPR17343] (ab179477)

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