Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-p53 antibody [E26] (ab32389)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E26] to p53
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-p53 antibody [E26]
    See all p53 primary antibodies

  • Description

    Rabbit monoclonal [E26] to p53

  • Host species

    Rabbit

  • Specificity

    The antibody should recognize human wild-type and mutant p53.

  • Tested Applications & Species

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

  • Immunogen

    Synthetic peptide within Human p53 aa 1-100. The exact sequence is proprietary.
    Database link: P04637

  • Positive control

    • Jurkat whole cell lysate (ab7899), A431 cells and human breast carcinoma. ICC/IF KO: Hap1 cells (Hap1-p53 KO used as a negative cell line)

  • 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 [E26] (ab32389)
    Western blot - Anti-p53 antibody [E26] (ab32389)

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: p53 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HEK293 whole cell lysate (20 µg)
    Lane 4: A431 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab32389 observed at 50 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab32389 was shown to specifically react with p53 in wild type HAP1 cells. No band was observed when p53 knockout samples were used. Wild-type and p53 knockout samples were subjected to SDS-PAGE. ab32389 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [E26] (ab32389)
    Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [E26] (ab32389)

    ab32389 staining p53 in wild-type Hap1 cells (top panel) and p53 knockout Hap1 cells (bottom panel). The cells were fixed with 100% methanol (5 min), 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 ab32389 at 1µg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution 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) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo color red).  Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a high-content analysis system (Perkin Elmer, Operetta CLS™).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody [E26] (ab32389)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody [E26] (ab32389)

    Immunohistochemical analysis of paraffin embedded human breast carcinoma showing nuclear localization of p53 using ab32389 at 1/100 dilution.

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

  • Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [E26] (ab32389)
    Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [E26] (ab32389)
    Immunofluorescent staining of A431 cells using ab32389 at 1/100 dilution.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody [E26] (ab32389)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody [E26] (ab32389) Image from Zaravinos A et al., PLoS One. 2011 Apr 4;6(4):e18135. Fig 5.; doi:10.1371/journal.pone.0018135; April 4, 2011, PLoS ONE 6(4): e18135.

    Immunohistochemical analysis of Human urinary bladder cancer tissue, staining p53 with ab32389 at 1/250 dilution. Staining was detected using DAB.

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

  • Western blot - Anti-p53 antibody [E26] (ab32389)
    Western blot - Anti-p53 antibody [E26] (ab32389)
    Anti-p53 antibody [E26] (ab32389) at 1/1000 dilution + Jurkat cell lysate

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

  • Anti-p53 antibody [E26] (ab32389)
    Anti-p53 antibody [E26] (ab32389)

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