Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)

Overview

  • Product nameAnti-p53 (phospho S46) antibody [EP42Y]
    See all p53 primary antibodies
  • Description
    Rabbit monoclonal [EP42Y] to p53 (phospho S46)
  • Tested applicationsSuitable for: WB, IP, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Human
  • Immunogen

    within Human p53 aa 1-100 (phospho S46). The exact sequence is proprietary.
    Database link: P04637

  • Positive control
    • IHC-P: Human ovarian adenocarcinoma. WB: HepG2 cell lysate treated with etoposide.
  • General notes

    A trial size is available to purchase for this antibody.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Alternative version available:
    Anti-p53 (phospho S46) antibody (HRP) [EP42Y] (ab202864)
    Anti-p53 (phospho S46) antibody (Alexa Fluor® 647) [EP42Y] (ab202865)

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab76242 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 - 1/2000. Predicted molecular weight: 53 kDa.
IP 1/40.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/500.
  • Application notesIs unsuitable for Flow Cyt.
  • Target

    • FunctionActs 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 specificityUbiquitous. 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 diseaseNote=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 similaritiesBelongs to the p53 family.
    • DomainThe 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 localizationCytoplasm; 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

    Anti-p53 (phospho S46) antibody [EP42Y] images

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

      Lane 1 : HEK293 cells were treated with Etoposide whole cell lysates with p53 phospho S46 peptide
      Lane 2 : HEK293 cells were treated with Etoposide whole cell lysates with p53 non-phospho peptide
      Lane 3 : HEK293 cells were treated with Etoposide whole cell lysates

      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 : 8 seconds

      Blocking buffer 5% NFDM/TBST

      Diluting buffer 5% NFDM/TBST

    • 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

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

      Lane 1 : HepG2 whole cell lysate - treated with etopside
      Lane 2 : HepG2 whole cell lysate - untreated

      Lysates/proteins at 10 µg per lane.

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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


      Exposure time : 3 minutes

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Immunohistochemical analysis of human ovarian adenocarcinoma using ab76242 at 1/100 dilution.
    • Immunocytochemistry/immunofluorescence staining of 4% PFA fixed; 0.1% Triton-X permeabilized A431 (human epidermoid carcinoma) cells with ab76242 at dilution of 1/500. The secondary antibody used was Alexa Fluor® 488; goat anti-rabbit IgG (ab150077) at a dilution of 1/1000. Nucleus was counter-stained with DAPI (blue). ab7291, a mouse anti-tubulin antibody (1/500) was used to stain tubulin along with ab150120 (AlexaFluor®594 goat anti-mouse secondary, 1/1000).

      Confocal image showed increased nuclear staining after ETP (etoposide; 25 μM, 5h) treatment on A431 cells. The LP treatment decreased the increased nuclear staining caused by ETP.

      ab179477 (1/500) was used as a Pan control for ab76242. The results showed nuclear staining on untreated, ETP and ETP+LP treated Hela cells.

    Protocols

    References for Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)

    This product has been referenced in:
    • Jiang S  et al. Piwil2 inhibits keratin 8 degradation through promoting p38-induced phosphorylation to resist Fas-mediated apoptosis. Mol Cell Biol 34:3928-38 (2014). Read more (PubMed: 25113562) »
    • Volkova M  et al. Activation of the aryl hydrocarbon receptor by doxorubicin mediates cytoprotective effects in the heart. Cardiovasc Res 90:305-14 (2011). WB ; Mouse . Read more (PubMed: 21233252) »

    See all 3 Publications for this product

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    Since mouse and rat p53 do not contain S46, I can replace this antibody ab76242 with a different antibody to another phospho-site of p53 that is present in mouse and rat. Please let me know which antibody would b...

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    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...

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