Validated using a knockout cell line

Anti-p53 antibody [pAb122] (ab90363)

Reviews (1)Q&A (2)References (6)

Overview

  • Product name

    Anti-p53 antibody [pAb122]
    See all p53 primary antibodies

  • Description

    Mouse monoclonal [pAb122] to p53

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IF, ICC, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Dog, Human

  • Immunogen

    Tissue, cells or virus corresponding to Mouse p53. SV40 transformed 3T3 cells
    Database link: P02340

  • Positive control

    • A431, Mouse and Rat brain tissue extracts. ICC/IF KO: Hap1 WT cells (Hap1-TP53 KO used as a negative cell line)

  • General notes

    Isotype: IgG2B, IgG1k

Properties

Applications

Our Abpromise guarantee covers the use of ab90363 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 1/250. Predicted molecular weight: 44 kDa.
Flow Cyt Use at an assay dependent concentration. PubMed: 2540940

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

 
ICC/IF 1/500 - 1/100.

Signal can be observed in cells fixed with either methanol or paraformaldehyde
ICC Use at an assay dependent concentration. PubMed: 2155427
IP Use at an assay dependent concentration. PubMed: 2155427

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

    see all

  • 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

  • Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [pAb122] (ab90363)
    Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [pAb122] (ab90363)

    ab90363 staining p53 in wild-type Hap1 cells (top panel) and p53 knockout Hap1 cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab90363 at 1/500 dilution and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 μg/ml (shown in pseudo color red).  Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a high-content analysis system (Perkin Elmer, Operetta CLS™).

  • Western blot - Anti-p53 antibody [pAb122] (ab90363)
    Western blot - Anti-p53 antibody [pAb122] (ab90363)This image is courtesy of an Abreview by Anke Rauch.
    All lanes : Anti-p53 antibody [pAb122] (ab90363) at 1/2000 dilution

    Lane 1 : HCT-116 cell lysate, untreated
    Lane 2 : HCT-116 cell lysate + 10 µM CPT-11 for 24 hrs
    Lane 3 : Negative control: HCT-116 cell lysate, untreated
    Lane 4 : Negative control: HCT-116 cell lysate + 10 µM CPT-11 for 24 hrs

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated rabbit anti-mouse polyclonal at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 44 kDa
    Additional bands at: 130 kDa (possible non-specific binding), 35 kDa (possible non-specific binding), 55 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 5 minutes


    Blocked with 5% milk for 1 hour at 21oC.

    See Abreview

  • Western blot - Anti-p53 antibody [pAb122] (ab90363)
    Western blot - Anti-p53 antibody [pAb122] (ab90363)
    All lanes : Anti-p53 antibody [pAb122] (ab90363) at 1/250 dilution

    Lane 1 : A431
    Lane 2 : Rat brain tissue extract
    Lane 3 : Mouse brain tissue extract

    Developed using the ECL technique.

    Predicted band size: 44 kDa

References

This product has been referenced in:

  • Cui C  et al. Hypoxia-inducible gene 2 promotes the immune escape of hepatocellular carcinoma from nature killer cells through the interleukin-10-STAT3 signaling pathway. J Exp Clin Cancer Res 38:229 (2019). Read more (PubMed: 31142329) »
  • Singh A  et al. Imprinting of cerebral cytochrome P450s in offsprings prenatally exposed to cypermethrin augments toxicity on rechallenge. Sci Rep 6:37426 (2016). WB ; Rat . Read more (PubMed: 27853314) »
See all 6 Publications for this product

Publishing research using ab90363? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Western blot abreview for Anti-p53 antibody [pAb122]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HCT-116 (Colorectal cancer cell line))
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
30 µg
Treatment
10 µM CPT-11 for 24 hrs
Specification
HCT-116 (Colorectal cancer cell line)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Read More

Anke Rauch

Verified customer

Submitted Aug 24 2016

Question

A user has problem with ab13847, ab90363, and ab5714.
You can see detailed protocols used, WB photos and user information in attached files.

Read More
Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that these antibodies are not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would like to offer some suggestions to help optimize the results.

ab90363 and ab5714
These antibodies are mouse monoclonals therefore, the secondary antibody should be an anti-mouse antibody.

For example:

Rabbit polyclonal Secondary Antibody to Mouse IgG - H&L, ab6709 (https://www.abcam.com/ab6709)

Goat polyclonal Secondary Antibody to Mouse IgG - H&L, ab6708 (https://www.abcam.com/ab6708)

Please also note, that ab90363 antibody should only be stored at 4ºC for 1-2 weeks. For optimum antibody activity, store at -20ºC.

ab13847
This antibody should also only be stored at 4ºC for 1-2 weeks. For optimum antibody activity, store at -20ºC or -80ºC.

You could try loading 20 ug of sample into each well rather than the 5 ug used.

Alternatively, if this does not improve the results, I could send out a free of charge replacement from a different lot or a different antibody. For example, ab2302 (https://www.abcam.com/ab2302) has given good results in western blot on human samples.

Should these suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

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Question

Dear Madam / Sir,

Two customer would like to test some antibodies on different tissues from written on the antibodies' dataheets.

one of the customers would like to test Anti-p53 antibody [PAb 240] (ab26) on cat paraffin tissues (IHC-P) and to have a free of charge antibody after testing (after sending protocol details, staining results and photos surely).

Customer information:

Another customer would like to test antibodies, "Caspase 3 antibody (ab7980), p53 antibody [pAb122] (ab90363), Bax antibody [6A7] (ab5714), Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (FITC), pre-adsorbed (ab97068)" on human frozen sections (IHC-Frozen).

Customer Information:


So, could you sent codes for the testing for each antibodies? After they have codes they will buy the antibodies.

Thank you.
--
Best regards,

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Answer

Expiration date: 18. October 2012

DISCOUNT CODEs:

ab26 cat : 0000

ab7980 IHC-Fr : 0000
ab90363 IHC-Fr : 000
ab5714 IHC-Fr : 000

ab97068 IHC-Fr : 00000

I am very pleased to hear your customer would like to accept our offer.

This code will give your customer 1 freeprimary antibody per codebefore the expiration date.

To redeem this offer, please submit an Abreview forthe untested species or applicationand include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests.

The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways:

1) Call to place your order and mention the code to our customer service department;

2) Include the code in your fax order;

3) Place your order on the web and enter the promotional code.

Any feedback that your customer can provide will be greatly appreciated, whether positive or negative. If you or your customer has any further questions, please do not hesitate to contact us. We look forward to receiving your customer's Abreview and wish your customer luck with their research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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