Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Mutant p53 antibody [Y5] (ab32049)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y5] to Mutant p53
  • Suitable for: ICC/IF, WB, IHC-P, Flow Cyt, IP
  • Reacts with: Human

Overview

  • Product name

    Anti-Mutant p53 antibody [Y5]
    See all Mutant p53 primary antibodies

  • Description

    Rabbit monoclonal [Y5] to Mutant p53

  • Host species

    Rabbit

  • Specificity

    ab32049 recognises human mutant forms of p53 but not human p53 wild type. ab32049 showed a negative signal on wildtype p53 cell lines (HepG2, A549, MCF-7) and a positive signal on mutant p53 cell lines (T47-D, Raji, A431) in WB.

  • Tested Applications & Species

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

  • Immunogen

    Synthetic peptide within Human Mutant p53 aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: P04637
    (Peptide available as ab204287)

  • General notes

    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.

Properties

Images

  • Western blot - Anti-Mutant p53 antibody [Y5] (ab32049)
    Western blot - Anti-Mutant p53 antibody [Y5] (ab32049)
    All lanes : Anti-Mutant p53 antibody [Y5] (ab32049) at 1/10000 dilution

    Lane 1 : A431 (Human epidermoid carcinoma epithelial cell), Whole cell lysate
    Lane 2 : A549 (Human lung carcinoma epithelial cell), Whole cell lysate
    Lane 3 : MCF-7 (Human breast adenocarcinoma epithelial cell), Whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 44 kDa


    Exposure time: 180 seconds


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

    Lane1: Mutant p53 cell lines

    Lane2-3: Wildtype p53 cell lines

    Exposure time: 180 seconds

    Observed MW: 50KDa

  • Western blot - Anti-Mutant p53 antibody [Y5] (ab32049)
    Western blot - Anti-Mutant p53 antibody [Y5] (ab32049)
    All lanes : Anti-Mutant p53 antibody [Y5] (ab32049) at 1/500 dilution

    Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell), Whole cell lysate
    Lane 2 : A549(Human lung carcinoma epithelial cell), Whole cell lysate
    Lane 3 : MCF-7(Human breast adenocarcinoma epithelial cell), Whole cell lysate
    Lane 4 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell), Whole cell lysate
    Lane 5 : MDA-MB-435(Human mammary gland ductal carcinoma melanocyte), Whole cell lysate
    Lane 6 : T-47D (Human ductal breast epithelial tumor epithelial cell), Whole cell lysate
    Lane 7 : Raji (Human Burkitt's lymphoma B lymphocyte), Whole cell lysate
    Lane 8 : Ramos (Human Burkitt's lymphoma B lymphocyte), Whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 44 kDa


    Exposure time: 60 seconds


    Blocking/Diluting Buffer and concentration: 5% NFDM/TBST

    Lane 1-4: Wildtype p53 cell lines

    Lane 5-8: Mutant p53 cell lines

    Observed MW: 50 kDa

  • Western blot - Anti-Mutant p53 antibody [Y5] (ab32049)
    Western blot - Anti-Mutant p53 antibody [Y5] (ab32049)
    Anti-Mutant p53 antibody [Y5] (ab32049) at 1/500 dilution + A431 cell lysate

    Predicted band size: 44 kDa
    Observed band size: 50 kDa
    why is the actual band size different from the predicted?

  • Immunocytochemistry/ Immunofluorescence - Anti-Mutant p53 antibody [Y5] (ab32049)
    Immunocytochemistry/ Immunofluorescence - Anti-Mutant p53 antibody [Y5] (ab32049)

    Confocal image showing nuclear staining on A431 cell line.

    Immunocytochemistry/Immunofluorescence analysis of A431(Human epidermoid carcinoma cell line) labelling Mutant p53 with ab32049 at 1/100. Cells were fixed with 4% Paraformaldehyde (20 minutes) and permeabilized with 0.1% Triton X-100 (5 minutes). ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counterstained with ab195889, anti- alpha tubulin antibody (1/200) using an Alexa Fluor® 594-conjugated microtubule marker as the secondary. Nuclei were counterstained with DAPI (blue).

    Secondary Only Control: PBS was used instead of the primary antibody as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)

    ab32049 showing positive staining in Urinary bladder carcinoma tissue.

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

  • Flow Cytometry - Anti-Mutant p53 antibody [Y5] (ab32049)
    Flow Cytometry - Anti-Mutant p53 antibody [Y5] (ab32049)

    Flow Cytometry analysis of A431(human epidermoid carcinoma) cells labeling Mutant p53 with unpurified ab32049 at 1/20 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)

    Immunohistochemistry (Paraffin-embedded sections) using ab32049 at a dilution of 1/50 and human skin cancer

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)

    ab32049 showing positive staining in Glioma tissue.

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)

    ab32049 showing positive staining in Gastric adenocarcinoma tissue.

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)

    ab32049 showing positive staining in Breast carcinoma tissue.

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)

    Immunohistochemical analysis of paraffin embedded normal Human breast tissue (negative control) labeling p53 with ab32049.

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)

    Immunohistochemical analysis of paraffin embedded normal Human uterus tissue (negative control) labeling p53 with ab32049.

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

  • OI-RD Scanning - Anti-Mutant p53 antibody [Y5] (ab32049)
    OI-RD Scanning - Anti-Mutant p53 antibody [Y5] (ab32049)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Anti-Mutant p53 antibody [Y5] (ab32049)
    Anti-Mutant p53 antibody [Y5] (ab32049)

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