Recombinant
RabMAb

Anti-p53 (phospho S392) antibody [EP155Y] (ab33889)

Overview

  • Product name
    Anti-p53 (phospho S392) antibody [EP155Y]
    See all p53 primary antibodies
  • Description
    Rabbit monoclonal [EP155Y] to p53 (phospho S392)
  • 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: WB, IHC-P, IPmore details
    Unsuitable for: ICC/IF
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human p53 (phospho S392). The exact sequence is proprietary.
    Database link: P04637

  • Positive control
    • WB: HEK293, A431, PC-12, RAW 264.7, HEK-293 and MCF7 cell lysates. IHC-P: Human prostate adenocarcinoma tissue. IP: 293T lysate.
  • General notes

     A trial size is available to purchase for this antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab33889 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/5000. Detects a band of approximately 53 kDa (predicted molecular weight: 44 kDa).
IHC-P 1/100 - 1/250.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

IP 1/20.

For unpurified use at 1/80. 

  • Application notes
    Is unsuitable for ICC/IF.
  • Target

    • Function
      Acts as a tumor suppressor in many tumor types; induces growth arrest or apoptosis depending on the physiological circumstances and cell type. Involved in cell cycle regulation as a trans-activator that acts to negatively regulate cell division by controlling a set of genes required for this process. One of the activated genes is an inhibitor of cyclin-dependent kinases. Apoptosis induction seems to be mediated either by stimulation of BAX and FAS antigen expression, or by repression of Bcl-2 expression. Implicated in Notch signaling cross-over. Isoform 2 enhances the transactivation activity of isoform 1 from some but not all TP53-inducible promoters. Isoform 4 suppresses transactivation activity and impairs growth suppression mediated by isoform 1. Isoform 7 inhibits isoform 1-mediated apoptosis.
    • Tissue specificity
      Ubiquitous. Isoforms are expressed in a wide range of normal tissues but in a tissue-dependent manner. Isoform 2 is expressed in most normal tissues but is not detected in brain, lung, prostate, muscle, fetal brain, spinal cord and fetal liver. Isoform 3 is expressed in most normal tissues but is not detected in lung, spleen, testis, fetal brain, spinal cord and fetal liver. Isoform 7 is expressed in most normal tissues but is not detected in prostate, uterus, skeletal muscle and breast. Isoform 8 is detected only in colon, bone marrow, testis, fetal brain and intestine. Isoform 9 is expressed in most normal tissues but is not detected in brain, heart, lung, fetal liver, salivary gland, breast or intestine.
    • Involvement in disease
      Note=TP53 is found in increased amounts in a wide variety of transformed cells. TP53 is frequently mutated or inactivated in about 60% of cancers. TP53 defects are found in Barrett metaplasia a condition in which the normally stratified squamous epithelium of the lower esophagus is replaced by a metaplastic columnar epithelium. The condition develops as a complication in approximately 10% of patients with chronic gastroesophageal reflux disease and predisposes to the development of esophageal adenocarcinoma.
      Defects in TP53 are a cause of esophageal cancer (ESCR) [MIM:133239].
      Defects in TP53 are a cause of Li-Fraumeni syndrome (LFS) [MIM:151623]. LFS is an autosomal dominant familial cancer syndrome that in its classic form is defined by the existence of a proband affected by a sarcoma before 45 years with a first degree relative affected by any tumor before 45 years and another first degree relative with any tumor before 45 years or a sarcoma at any age. Other clinical definitions for LFS have been proposed (PubMed:8118819 and PubMed:8718514) and called Li-Fraumeni like syndrome (LFL). In these families affected relatives develop a diverse set of malignancies at unusually early ages. Four types of cancers account for 80% of tumors occurring in TP53 germline mutation carriers: breast cancers, soft tissue and bone sarcomas, brain tumors (astrocytomas) and adrenocortical carcinomas. Less frequent tumors include choroid plexus carcinoma or papilloma before the age of 15, rhabdomyosarcoma before the age of 5, leukemia, Wilms tumor, malignant phyllodes tumor, colorectal and gastric cancers.
      Defects in TP53 are involved in head and neck squamous cell carcinomas (HNSCC) [MIM:275355]; also known as squamous cell carcinoma of the head and neck.
      Defects in TP53 are a cause of lung cancer (LNCR) [MIM:211980].
      Defects in TP53 are a cause of choroid plexus papilloma (CPLPA) [MIM:260500]. Choroid plexus papilloma is a slow-growing benign tumor of the choroid plexus that often invades the leptomeninges. In children it is usually in a lateral ventricle but in adults it is more often in the fourth ventricle. Hydrocephalus is common, either from obstruction or from tumor secretion of cerebrospinal fluid. If it undergoes malignant transformation it is called a choroid plexus carcinoma. Primary choroid plexus tumors are rare and usually occur in early childhood.
      Defects in TP53 are a cause of adrenocortical carcinoma (ADCC) [MIM:202300]. ADCC is a rare childhood tumor of the adrenal cortex. It occurs with increased frequency in patients with the Beckwith-Wiedemann syndrome and is a component tumor in Li-Fraumeni syndrome.
    • Sequence similarities
      Belongs to the p53 family.
    • Domain
      The nuclear export signal acts as a transcriptional repression domain. The TADI and TADII motifs (residues 17 to 25 and 48 to 56) correspond both to 9aaTAD motifs which are transactivation domains present in a large number of yeast and animal transcription factors.
    • Post-translational
      modifications
      Acetylated. Acetylation of Lys-382 by CREBBP enhances transcriptional activity. Deacetylation of Lys-382 by SIRT1 impairs its ability to induce proapoptotic program and modulate cell senescence.
      Phosphorylation on Ser residues mediates transcriptional activation. Phosphorylated by HIPK1 (By similarity). Phosphorylation at Ser-9 by HIPK4 increases repression activity on BIRC5 promoter. Phosphorylated on Thr-18 by VRK1. Phosphorylated on Ser-20 by CHEK2 in response to DNA damage, which prevents ubiquitination by MDM2. Phosphorylated on Thr-55 by TAF1, which promotes MDM2-mediated degradation. Phosphorylated on Ser-46 by HIPK2 upon UV irradiation. Phosphorylation on Ser-46 is required for acetylation by CREBBP. Phosphorylated on Ser-392 following UV but not gamma irradiation. Phosphorylated upon DNA damage, probably by ATM or ATR. Phosphorylated on Ser-15 upon ultraviolet irradiation; which is enhanced by interaction with BANP.
      Dephosphorylated by PP2A-PPP2R5C holoenzyme at Thr-55. SV40 small T antigen inhibits the dephosphorylation by the AC form of PP2A.
      May be O-glycosylated in the C-terminal basic region. Studied in EB-1 cell line.
      Ubiquitinated by MDM2 and SYVN1, which leads to proteasomal degradation. Ubiquitinated by RFWD3, which works in cooperation with MDM2 and may catalyze the formation of short polyubiquitin chains on p53/TP53 that are not targeted to the proteasome. Ubiquitinated by MKRN1 at Lys-291 and Lys-292, which leads to proteasomal degradation. Deubiquitinated by USP10, leading to its stabilization. Ubiquitinated by TRIM24, which leads to proteasomal degradation. Ubiquitination by TOPORS induces degradation. Deubiquitination by USP7, leading to stabilization. Isoform 4 is monoubiquitinated in an MDM2-independent manner.
      Monomethylated at Lys-372 by SETD7, leading to stabilization and increased transcriptional activation. Monomethylated at Lys-370 by SMYD2, leading to decreased DNA-binding activity and subsequent transcriptional regulation activity. Lys-372 monomethylation prevents interaction with SMYD2 and subsequent monomethylation at Lys-370. Dimethylated at Lys-373 by EHMT1 and EHMT2. Monomethylated at Lys-382 by SETD8, promoting interaction with L3MBTL1 and leading to repress transcriptional activity. Demethylation of dimethylated Lys-370 by KDM1A prevents interaction with TP53BP1 and represses TP53-mediated transcriptional activation.
      Sumoylated by SUMO1.
    • Cellular localization
      Cytoplasm; Cytoplasm. Nucleus. Nucleus > PML body. Endoplasmic reticulum. Interaction with BANP promotes nuclear localization. Recruited into PML bodies together with CHEK2; Nucleus. Cytoplasm. Localized in both nucleus and cytoplasm in most cells. In some cells, forms foci in the nucleus that are different from nucleoli; Nucleus. Cytoplasm. Localized in the nucleus in most cells but found in the cytoplasm in some cells; Nucleus. Cytoplasm. Localized mainly in the nucleus with minor staining in the cytoplasm; Nucleus. Cytoplasm. Predominantly nuclear but localizes to the cytoplasm when expressed with isoform 4 and Nucleus. Cytoplasm. Predominantly nuclear but translocates to the cytoplasm following cell stress.
    • Information by UniProt
    • Database links
    • Alternative names
      • Antigen NY-CO-13 antibody
      • BCC7 antibody
      • Cellular tumor antigen p53 antibody
      • FLJ92943 antibody
      • LFS1 antibody
      • Mutant tumor protein 53 antibody
      • p53 antibody
      • p53 tumor suppressor antibody
      • P53_HUMAN antibody
      • Phosphoprotein p53 antibody
      • Tp53 antibody
      • Transformation related protein 53 antibody
      • TRP53 antibody
      • Tumor protein 53 antibody
      • Tumor protein p53 antibody
      • Tumor suppressor p53 antibody
      see all

    Images

    • 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.
    • 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



      Blocking and diluting buffer: 5% NFDM/TBST.

    • 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



      Blocking and diluting buffer: 5% NFDM/TBST.

    • 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?)



      Blocking and diluting buffer: 5% NFDM/TBST

    • 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 secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1:1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    • 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


      Blocking and dilution buffer: 5% NFDM/TBST.

    • Unpurified ab33889, at a 1/100 dilution, staining p53 in paraffin embedded human prostate adenocarcinoma tissue by Immunohistochemistry.

    • 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.

    • All lanes : Anti-p53 (phospho S392) antibody [EP155Y] (ab33889) at 1/1000 dilution (unpurified)

      Lane 1 : HEK293 whole cell lysate - untreated
      Lane 2 : HEK293 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


      Blocking and dilution buffer: 5% NFDM/TBST.

    • 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.

    • 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?)

    References

    This product has been referenced in:
    • Kotipatruni RR  et al. p53- and Bax-mediated apoptosis in injured rat spinal cord. Neurochem Res 36:2063-74 (2011). WB, IHC (PFA fixed) ; Rat . Read more (PubMed: 21748659) »

    See 1 Publication for this product

    Customer reviews and Q&As

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.5% Triton X-100 in PBS
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Username

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Jul 03 2018

    Thank you for your patience.
    I am happy to confirm that the cells have been treated with 5 ug/ml Actinomycin D for 3 hrs, 6 hrs and 18 hrs.
    I hope this information is helpful. Please do not hesitate to contact us again, should you need any ad...

    Read More

    Thank you for your call yesterday.
    I can confirm that for the positive control for ab76242 the HepG2 cells are treated with 25uM etoposide for 5 hours. I'd like to mention again though, that the laboratory confirmed that the flow cytometry results...

    Read More

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