Overview

  • Product name
    p53 Antibody Sampler Panel
  • Product overview

    ab219089 is a sampler panel consisting of thoroughly-validated recombinant monoclonal antibodies against several variants of p53. This panel includes trial sizes of antibodies to unmodified p53, mutant p53, p53 (acetylated Lysine 382), p53 (phosphorylated Serine 20), p53 (phosphorylated Serine 392), p53 (phosphorylated Serine 46), and two anti-rabbit IgG (HRP) and anti-mouse IgG (HRP) antibodies.

Properties

Images

  • Predicted band size : 43.6 kDa

    Lanes 1, 5 and 9: Wild-type HAP1 cell lysate (20 µg)
    Lanes 2, 6 and 10: p53 knockout HAP1 cell lysate (20 µg)
    Lanes 3, 7 and 11: A431 cell lysate (20 µg)
    Lanes 4, 8 and 12: Saos-2 cell lysate (20 µg)
    Lanes 1, 2, 3 and 4: Green signal from target – ab1101 observed at 53 kDa
    Lanes 5, 6, 7 and 8: Red signal from loading control – ab181602 observed at 37 kDa
    Lanes 9, 10, 11 and 12: Merged (red and green) signal

    ab1101 was shown to specifically react with p53 when p53 knockout samples were used. Wild-type and p53 knockout samples were subjected to SDS-PAGE. ab1101 and ab181602 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4ºC. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • ab32049 showing positive staining in urinary bladder carcinoma tissue.

  • All lanes : Anti-p53 (phospho S20) antibody [EPR2156(2)] (ab157454) at 1/1000 dilution

    Lane 1 : T47D (human mammary gland ductal carcinoma) treated with Etoposide whole cell lysate
    Lane 2 : Untreated T47D (human mammary gland ductal carcinoma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution

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


    Exposure time : 1 minute

    Blocking & dilution buffer: 5% NFDM/TBST

  • IHC image of ab75754 staining p53 (acetyl K382) in human breast adenocarcinoma formalin fixed paraffin embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab75754, 1μg/ml working concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • All lanes : Anti-p53 (phospho S46) antibody [EP42Y] (ab76242) at 1/5000 dilution

    Lane 1 : Untreated HepG2 whole cell lysate
    Lane 2 : HepG2 whole cell lysate, treated with etoposide
    Lane 3 : HepG2 whole cell lysate, treated with etoposide, then the membrane was treated with alkaline phosphatase

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 53 kDa
    Observed band size : 53 kDa


    Exposure time : 1 minute

    Blocking buffer: 2% BSA/TBST
    Dilution buffer: 2% BSA/TBST

  • Ab33889, at a 1/100 dilution, staining p53 in paraffin embedded human prostate adenocarcinoma tissue by Immunohistochemistry.

References

ab219089 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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