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RecombinantRabMAb

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

Submit a review Q&A (5)References (9)
Western blot - Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)
  • Immunocytochemistry/ Immunofluorescence - Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)
  • Western blot - Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)
  • Western blot - Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)
  • Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP42Y] to p53 (phospho S46)
  • Suitable for: WB, IP, IHC-P, ICC/IF
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free HRP

Overview

  • Product name

    Anti-p53 (phospho S46) antibody [EP42Y]
    See all p53 primary antibodies

  • Description

    Rabbit monoclonal [EP42Y] to p53 (phospho S46)

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IP, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

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

  • General notes

    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.

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

Properties

Applications

The Abpromise guarantee

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.
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 notes
Is unsuitable for Flow Cyt.

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 antigen p55 antibody
      • Tumor protein 53 antibody
      • Tumor protein p53 antibody
      • Tumor suppressor p53 antibody
      see all

    Images

    • Western blot - Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)
      Western blot - Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)
      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
      All lanes : 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)

      Immunohistochemical analysis of human ovarian adenocarcinoma using ab76242 at 1/100 dilution.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)
      Immunocytochemistry/ Immunofluorescence - Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)

      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.

    • Western blot - Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)
      Western blot - Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)
      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
      All lanes : 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

    • Western blot - Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)
      Western blot - Anti-p53 (phospho S46) antibody [EP42Y] (ab76242)
      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
      All lanes : 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.

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

    References (9)

    Publishing research using ab76242? Please let us know so that we can cite the reference in this datasheet.

    ab76242 has been referenced in 9 publications.

    • Sari C  et al. Caffeic acid phenethyl ester induces apoptosis in colorectal cancer cells via inhibition of survivin. Turk J Biol 44:264-274 (2020). PubMed: 33110364
    • Cheng J  et al. Inhibition of invasive pancreatic cancer: restoring cell apoptosis by activating mitochondrial p53. Am J Cancer Res 9:390-405 (2019). PubMed: 30906636
    • Yang R  et al. Cell type-dependent bimodal p53 activation engenders a dynamic mechanism of chemoresistance. Sci Adv 4:eaat5077 (2018). PubMed: 30585287
    • Liu L  et al. Variant 2 of KIAA0101, antagonizing its oncogenic variant 1, might be a potential therapeutic strategy in hepatocellular carcinoma. Oncotarget 8:43990-44003 (2017). PubMed: 28410205
    • Cheng ZX  et al. MicroRNA-132 induces temozolomide resistance and promotes the formation of cancer stem cell phenotypes by targeting tumor suppressor candidate 3 in glioblastoma. Int J Mol Med 40:1307-1314 (2017). PubMed: 28901390
    • 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). PubMed: 25113562
    • De S  et al. RECQL4 is essential for the transport of p53 to mitochondria in normal human cells in the absence of exogenous stress. J Cell Sci 125:2509-22 (2012). PubMed: 22357944
    • 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 . PubMed: 21233252
    • Idikio HA Immunohistochemistry in diagnostic surgical pathology: contributions of protein life-cycle, use of evidence-based methods and data normalization on interpretation of immunohistochemical stains. Int J Clin Exp Pathol 3:169-76 (2009). PubMed: 20126585

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-5 of 5 Abreviews or Q&A

    Question

    Please can we have ab13533 - Anti-SOD2 antibody?

    Read More
    Answer

    Thank you for your reply.

    I am sorry this product did not work in mouse and rat as previously stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab13533.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More

    Question

    I bought the antibody ab76242. We only work on mouse cells and rodent
    tissues and we got negative results. But without positive controls, it
    was hard for us to incriminate the reactivity of the antibody.
    So please find enclosed the bill corresponding to our purchase. We benefited from a discount offer, I hope it will fine for the refund.
    Thank you for your help.

    Read More
    Answer

    Thank you for contacting us.

    Your credit note ID is xxx.

    I am sorry that this antibody had incorrect information on the datasheet - which we have corrected in the meantime. I have asked our Finance department to issue a credit note for you which is for the original price of the antibody. The credit note may be used in one of the following ways:
    (1) Redeemed against the original invoice if this hasn't already been paid.
    (2) Held on the account for use against a future order.
    (3) A full refund can be offered where no other invoices are outstanding.
    Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.
    To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.
    The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.
    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice.

    Read More

    Question

    I really appreciate your update information and compensatory offer for the problem. I ordered your product ab76242. We were and is still working on rat or mouse neurons and we didn't get any significant signal in our Western blotting with the antibody, where as your another p53 antibody ab28, we bought around that time, worked fine. Since the antibody, ab76242, was useless for our work, I want to get an replacement with your other anti-p53 antibody [DO-1], ab80645, which is reported to work on mouse p53. We really appreciate your response as soon as possible, since we have already wasted our time significantly.

    Read More
    Answer

    Thank you for your reply.

    I am sorry that we had incorrect information about this product on our datasheet - which we have corrected by now -and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab80645.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.


    I wish you the best of luck with your research.

    Read More

    Question

    We only study mouse / rat and would explain this not working for us. Whatt shoud we do?

    Read More
    Answer

    Thank you for contacting us.

    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 be best or let me know the phospho-site and I can search our catalog for an according antibody.

    Alternatively, a credit or refund can be arranged.

    I apologize for the inconvience, butwould be happy to resolve this case to your satisfaction.

    I look forward to your reply.

    Use our products? Submit an Abreview. Earn rewards!
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    Question

    phone call: how where the cells in the positive controlstimulated - for how long?

    Read More
    Answer

    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 were negative, so we do not recommend this application.
    Unfortunately the lab could not yet confirm the stimulation protocol details for the ab33889. I will forward it to you, as soon as I receive the answer.
    Please do not hesitate to contact us also meanwhile, if you have any further question.

    Read More

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