Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP155Y] to p53 (phospho S392) - BSA and Azide free
  • Suitable for: IHC-P, WB, Dot blot, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free
    See all p53 primary antibodies

  • Description

    Rabbit monoclonal [EP155Y] to p53 (phospho S392) - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    This antibody is specific for p53 phosphorylated on Serine 392.  The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Tested applications

    Suitable for: IHC-P, WB, Dot blot, IPmore details
    Unsuitable for: ICC/IF

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • General notes

    Ab239211 is the carrier-free version of ab33889. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab239211 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    All lanes : Anti-p53 (phospho S392) antibody [EP155Y] (ab33889) at 1/1000 dilution (purified)

    Lane 1 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
    Lane 2 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates, then the membrane was incubated with alkaline phosphatase.

    Lysates/proteins at 15 µg per lane.

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

    Predicted band size: 44 kDa



    This data was developed using ab33889, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

    This data was developed using ab33889, the same antibody clone in a different buffer formulation.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling p53 with Purified ab33889 at 1:250 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
  • Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    All lanes : Anti-p53 (phospho S392) antibody [EP155Y] (ab33889) at 1/1000 dilution (purified)

    Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
    Lane 2 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates, then the membrane was incubated with alkaline phosphatase.

    Lysates/proteins at 15 µg per lane.

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

    Predicted band size: 44 kDa



    This data was developed using ab33889, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    All lanes : Anti-p53 (phospho S392) antibody [EP155Y] (ab33889) at 1/1000 dilution (purified)

    Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates, then the membrane was incubated with alkaline phosphatase.

    Lysates/proteins at 15 µg per lane.

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

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



    This data was developed using ab33889, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunoprecipitation - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    Immunoprecipitation - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

    This data was developed using ab33889, the same antibody clone in a different buffer formulation.

    ab33889 (purified) at 1:20 dilution (0.6μg) immunoprecipitating p53 in 293T whole cell lysate.

    Lane 1 (input): 293T (Human embryonic kidney epithelial cell) whole cell lysate, 10μg
    Lane 2 (+): ab33889 & 293T whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab33889 in 293T whole cell lysate.

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.
  • Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    All lanes : Anti-p53 (phospho S392) antibody [EP155Y] (ab33889) at 1/2000 dilution (purified)

    Lane 1 : A431 whole cell lysate
    Lane 2 : A431 treated with 1μg/ml doxorubicin for 24 hours whole cell lysate
    Lane 3 : A431 treated with 1μg/ml doxorubicin for 24 hours whole cell lysate, the membrane was incubated with alkaline phosphatase.

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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


    Exposure time: 15 seconds


    This data was developed using ab33889, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

    This data was developed using ab33889, the same antibody clone in a different buffer formulation.

    Unpurified ab33889, at a 1/100 dilution, staining p53 in paraffin embedded human prostate adenocarcinoma tissue by Immunohistochemistry.
  • Dot Blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    Dot Blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    This data was developed using ab33889, the same antibody clone in a different buffer formulation.Dot blot analysis of p53 (pS392) peptide (Lane 1) and p53 non-phospho peptide (Lane 2) labelling p53 (pS392) with unpurified ab33889 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000. Blocking and dilution buffer: 5% NFDM/TBST. Exposure time: 3 minutes.
  • Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    All lanes : Anti-p53 (phospho S392) antibody [EP155Y] (ab33889) at 1/1000 dilution (unpurified)

    Lane 1 : HEK-293 whole cell lysate - untreated
    Lane 2 : HEK-293 whole cell lysate - treated with Alkaline Phosphatase

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG, (H+L) 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: 15 seconds


    This data was developed using ab33889, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

    This data was developed using ab33889, the same antibody clone in a different buffer formulation.

    Lanes 1 and 5: Extract of Hek293T incubated with etoposide (7.5 µg).

    Lanes 2 and 6: Lambda phosphatase (400 times-diluted)-treated extract of Hek293T incubated with etoposide (7.5 µg).

    Lanes 3 and 7:  Lambda phosphatase (100 times-diluted)-treated extract of Hek293T incubated with etoposide (7.5 µg).

    Lanes 4 and 8: Lambda phosphatase (25 times-diluted)-treated extract of Hek293T incubated with etoposide (7.5 µg).

    SDS PAGE performed under reducing conditions (100mM DTT, sample heated at 50°C).

    Primary:

    Lanes 1-4: Anti p53 (phosphoS392) antibody (ab33889, unpurified) at 1/2000 dilution. 

    Lanes 5-8: Anti p53 antibody (ab1101) at 1/2500 dilution.

    Secondary:

    Lanes 1-4: HRP-conjugated goat anti-rabbit IgG (H&L) at 1/10000.

    Lanes 5-8: HRP-conjugated goat anti-mouse IgG (H&L) at 1/10000.

    Blocked in 5% milk in PBS for 3 hours at room temperature.

    Incubated with the primary antibody in 5% BSA + 50mM Tris pH 7.5 + 0.05% Tween-20 overnight at 4°C.

    Incubated with the secondary antibody in blocking buffer for 2 hours at room temperature.

  • Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    All lanes : Anti-p53 (phospho S392) antibody [EP155Y] (ab33889) at 1/500 dilution (unpurified)

    Lane 1 : MCF7 cell lysate untreated
    Lane 2 : MCF7 cell lysate treated with 5 ug/ml Actinomycin for 3hrs.
    Lane 3 : MCF7 cell lysate treated with 5 ug/ml Actinomycin for 6hrs.
    Lane 4 : MCF7 cell lysate treated with 5 ug/ml Actinomycin for 18hrs.

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



    This data was developed using ab33889, the same antibody clone in a different buffer formulation.

  • Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)
    Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

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