Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-p53 antibody [PAb 240] - BSA and Azide free (ab176243)

Key features and details

  • Mouse monoclonal [PAb 240] to p53 - BSA and Azide free
  • Suitable for: ICC/IF, WB, IP
  • Reacts with: Mouse, Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-p53 antibody [PAb 240] - BSA and Azide free
    See all p53 primary antibodies

  • Description

    Mouse monoclonal [PAb 240] to p53 - BSA and Azide free

  • Host species

    Mouse

  • Tested applications

    Suitable for: ICC/IF, WB, IPmore details

  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow, Dog, Syrian hamster

  • Immunogen

    Fusion protein corresponding to Mouse p53 aa 1-300. Gel-purified p53-beta-galactosidase fusion protein containing murine p53 from aa 14-389 (derived from pSV53C cDNA clone).
    Database link: P02340

  • Epitope

    The epitope has been mapped between amino acids 213 and 217 on human p53.

  • Positive control

    • ICC/IF: A431 cells. IP: HCT116 whole cell lysate WB: NIH/3T3 cells treated with 1 µM doxorubicin for 24 hours lysate, HeLa cells untreated and Bleomycin treated lysates, MCF7 cells (p53 WT and mutant) whole cell lysate.

  • General notes

    This product is the BSA- and Azide-Free version of Mouse Monoclonal [PAb 240] to p53 (ab26). 

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-p53 antibody [PAb 240] - BSA and Azide free (ab176243)
    Western blot - Anti-p53 antibody [PAb 240] - BSA and Azide free (ab176243)
    All lanes : Anti-p53 antibody [PAb 240] (ab26) at 5 µg/ml

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

    Performed under reducing conditions.

    Predicted band size: 43 kDa
    Observed band size: 53 kDa
    why is the actual band size different from the predicted?



    Lanes 1-6: Merged (red and green) signal.


    Ab26 was shown to specifically react with p53 in wild type HCT116 cells treated with irinotecan. No band was observed in p53 knockout HCT116 cells. Wild-type and p53 knockout samples, positive and negative controls were subjected to SDS-PAGE. Ab26 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.

    Wild-type and p53 knockout HCT116 cell lysates were kindly provided by a collaborator.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-Arginine and sodium azide (ab26).

  • Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [PAb 240] - BSA and Azide free (ab176243)
    Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [PAb 240] - BSA and Azide free (ab176243)

    ab26 stained in A431 cells. Cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with ab26 at 1µg/ml and ab6046 (Rabbit polyclonal to beta tubulin) at 1ug/ml overnight at +4°C. The secondary antibodies were ab150177 (colored green) used at 1 ug/ml and ab150087 (pseudo-colored red) used at 2ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-Arginine and sodium azide (ab26).

  • Immunoprecipitation - Anti-p53 antibody [PAb 240] - BSA and Azide free (ab176243)
    Immunoprecipitation - Anti-p53 antibody [PAb 240] - BSA and Azide free (ab176243)

    p53 was immunoprecipitated from 7x106 HCT116 (human colon carcinoma cell line) cells with ab26 at 1/150 dilution. Western blot was performed from the immunoprecipitate using anti-p53 antibody. Donkey Anti-Mouse IgG H&L (Alexa Fluor® 750) preadsorbed (ab175739) was used as secondary antibody at 1/5000 dilution.


    Lane 1: HCT116 whole cell lysate 10 µg (Input).

    Lane 2: ab207799 IP in etoposide treated HCT116 whole cell lysate.

    Lane 3: ab207799 IP in etoposide treated HCT116 p53-/- whole cell lysate (negative control).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-Arginine and sodium azide (ab26).

  • Western blot - Anti-p53 antibody [PAb 240] - BSA and Azide free (ab176243)
    Western blot - Anti-p53 antibody [PAb 240] - BSA and Azide free (ab176243)

    Primary: All Lanes: Anti-p53 antibody (ab26) at 5 µg/mL. Lane 1: MW marker. Lane 2: NIH/3T3 cells treated with vehicle for 24 hours. Lane 3: NIH/3T3 cells treated with 1 µM doxorubicin for 24 hours Secondary: All Lanes: HRP-conjugated VeriBlot anti-Mouse IgG (ab131368) 1:1000. Lysates at 20 µg/lane. Performed under denaturing conditions. Developed using ECL technique. Blocking buffer: 5% milk in PBS.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-Arginine and sodium azide (ab26).

  • Western blot - Anti-p53 antibody [PAb 240] - BSA and Azide free (ab176243)
    Western blot - Anti-p53 antibody [PAb 240] - BSA and Azide free (ab176243)
    Anti-p53 antibody [PAb 240] (ab26) at 1 µg/ml + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate, 10 ug

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 43 kDa


    Exposure time: 8 minutes


    This blot was produced using a 4-12% 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 2% Bovine Serum Albumin before being incubated with ab26 overnight at 4°C. Antibody binding was detected using an anti-mouse HRP secondary antibody, and visualised using ECL development solution ab133406.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-Arginine and sodium azide (ab26).

  • Western blot - Anti-p53 antibody [PAb 240] - BSA and Azide free (ab176243)
    Western blot - Anti-p53 antibody [PAb 240] - BSA and Azide free (ab176243)
    All lanes : Anti-p53 antibody [PAb 240] (ab26) at 5 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Hela Whole Cell Lysate - Bleomycin Treated (40U/ml),

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 43 kDa


    Exposure time: 4 minutes


    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-Arginine and sodium azide (ab26).

  • Western blot - Anti-p53 antibody [PAb 240] - BSA and Azide free (ab176243)
    Western blot - Anti-p53 antibody [PAb 240] - BSA and Azide free (ab176243)
    All lanes : Anti-p53 antibody [PAb 240] (ab26) at 1 µg/ml

    All lanes : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP), at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 43 kDa


    Exposure time: 4 minutes


    Lanes 1-2: 1% BSA blocking buffer

    Lanes 3-4: 3% Milk blocking buffer

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-Arginine and sodium azide (ab26).

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