Overview

  • Product name
    Anti-p53 antibody [PAb 240]
    See all p53 primary antibodies
  • Description
    Mouse monoclonal [PAb 240] to p53
  • Specificity
    This p53 monoclonal antibody recognizes both mutant and wild type conformational forms of p53 under denaturing conditions. The PAb 240 clone recognizes an epitope that is structurally hidden in the wild type conformation of p53 but becomes exposed by denaturation or in mutant conformations of p53 where point mutations in the TP53 gene alter the structure of the protein (Gannon JV et al., 1990; Stephen CW et al., 1992; Wang PL et al., 2001).
  • Tested applications
    Suitable for: IP, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Dog, Human, Monkey, Chinese hamster, Syrian hamster
  • Immunogen

    Gel-purified p53-beta-galactosidase fusion protein containing murine p53 from aa 14-389 (derived from pSV53C cDNA clone).

  • Epitope
    The epitope has been mapped between amino acids 213 and 217 on human and mouse p53.
  • Positive control
    • WB: A431 cell lysate. ICC-IF: A431 cells.
  • General notes

    This p53 antibody has been knockout validated in Western blot. The expected band was seen in wild type HCT116 cells treated with the DNA damaging agent irinotecan and no band was seen in TP53 knockout HCT116 cells.

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

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab26 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
IP Use a concentration of 10 µg/ml.
ICC/IF Use a concentration of 0.5 - 1 µg/ml.
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).

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

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

  • All lanes : Anti-p53 antibody [PAb 240] (ab26) at 5 µg/ml

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

    Developed using the ECL technique

    Performed under reducing conditions.

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

    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.

  • 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.



  • Predicted band size : 53 kDa

    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.

  • Lanes 1 - 2 : Anti-p53 antibody [PAb 240] (ab26) at 1 µg/ml
    Lanes 3 - 4 : Anti-p53 antibody [PAb 240] (ab26) at 5 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    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 : 53 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.

  • 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
    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 : 53 kDa


    Exposure time : 4 minutes
  • Anti-p53 antibody [PAb 240] (ab26) at 1 µg/ml + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

    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 : 53 kDa
    Observed band size : 53 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

  • All lanes : Anti-p53 antibody [PAb 240] (ab26) at 1/2000 dilution

    Lane 1 : Human breast cancer cell-line, MCF7 cells (p53 WT), whole cell lysate
    Lane 2 : Human breast cancer cell-line, MDA231 cells (p53 Mutant), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP conjugated donkey anti-mouse antibody at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 53 kDa
    Observed band size : 53 kDa
    Additional bands at : 72 kDa (possible non-specific binding).

    Exposure time : 10 seconds

    This image is courtesy of an Abreview submitted by Dr Cherie Blenkiron

    See Abreview

  • 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).

References

This product has been referenced in:
  • Singh A  et al. Podophyllotoxin and Rutin Modulates Ionizing Radiation-Induced Oxidative Stress and Apoptotic Cell Death in Mice Bone Marrow and Spleen. Front Immunol 8:183 (2017). Read more (PubMed: 28289414) »
  • Fan L  et al. PADI4 Epigenetically Suppresses p21 Transcription and Inhibits Cell Apoptosis in Fibroblast-like Synoviocytes from Rheumatoid Arthritis Patients. Int J Biol Sci 13:358-366 (2017). WB ; Human . Read more (PubMed: 28367100) »

See all 82 Publications for this product

Customer reviews and Q&As

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The one antibody that I see mentioned most often in an online search as mutant p53-specific is the clone PAb 240. The following paper describes the production and characterization of the clone, which Abcam carrie...

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