製品の概要

  • 製品名Anti-p53 antibody [DO-1]
    p53 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [DO-1] to p53
  • 特異性This antibody recognizes an epitope within aa 20-25 at the N-terminus of human p53.
  • アプリケーション適用あり: ChIP, ICC/IF, ELISA, IHC-P, IHC-Fr, IP, WB, Flow Cytmore details
  • 種交差性
    交差種: Human
    非交差種: Mouse, Rat
  • 免疫原

    Recombinant full length protein corresponding to Human p53 (N terminal). epitope is within aa 20-25
    Database link: P04637

  • ポジティブ・コントロール
    • The antibody gave a positive signal in Human Brain tissue as well as the following whole cell lysates: HBL 100 cells, Human HepG2, MCF7, HEK293, DU145. IHC-P - Human colon adenocarcinoma FFPE tissue sections
  • 特記事項

    Dilute in PBS or medium which is identical to that used in the assay system.

    Alternative versions available:

    Anti-p53 antibody (Biotin) [DO-1] (ab27696)
    Anti-p53 antibody (Phycoerythrin) [DO-1] (ab27697)
    Anti-p53 antibody (Alexa Fluor® 488) [DO-1] (ab204245)

    Anti-p53 antibody (HRP) [DO-1] (ab204452)

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab1101 in the following tested applications.

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

アプリケーション Abreviews 特記事項
ChIP Use at an assay dependent concentration.

Use at an assay dependent concentration.

ICC/IF Use at an assay dependent concentration.
ELISA Use a concentration of 1 - 5 µg/ml.
IHC-P Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr 1/100.
IP Use a concentration of 1 - 5 µg/ml.
WB 1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 43.6 kDa).
Flow Cyt 1/200.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

ターゲット情報

  • 機能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.
  • 組織特異性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.
  • 関連疾患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.
  • 配列類似性Belongs to the p53 family.
  • ドメイン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.
  • 翻訳後修飾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.
  • 細胞内局在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
  • 参照データベース
  • 別名
    • 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 antibody [DO-1] 画像



  • Predicted band size : 43.6 kDa

    Lanes 1, 5 and 9: Wild-type HAP1 cell lysate (20 µg)
    Lanes 2, 6 and 10: p53 knockout HAP1 cell lysate (20 µg)
    Lanes 3, 7 and 11: A431 cell lysate (20 µg)
    Lanes 4, 8 and 12: Saos-2 cell lysate (20 µg)
    Lanes 1, 2, 3 and 4: Green signal from target – ab1101 observed at 53 kDa
    Lanes 5, 6, 7 and 8: Red signal from loading control – ab181602 observed at 37 kDa
    Lanes 9, 10, 11 and 12: Merged (red and green) signal

    ab1101 was shown to specifically react with p53 when p53 knockout samples were used. Wild-type and p53 knockout samples were subjected to SDS-PAGE. ab1101 and ab181602 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4ºC. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) ab216772 and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) ab216777 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • IHC image of ab1101 staining p53 in human colon adenocarcinoma formalin-fixed paraffin-embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab1101, 1/500 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. High magnification of the tumor region - T (lower right panel) and adjacent normal crypts - N (lower left panel) are shown.
    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Chromatin was prepared from 293 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab1101, and 10ml of protein A sepharose beads,10ml of protein G sepharose beads. No IgG was added to the beads control. The immunoprecipitated DNA was quantified by real time PCR. Primers were as follows:

    Bax-1, forward: GGGTTATCTCTTGGGCTCACAA.

    Bax-1, reverse: GAGCTCTCCCCAGCGCA.

    Bax-2, forward: TGG AGC TGC AGA GGA TGA TTG

    Bax-2, reverse: CCA GTT GAA GTT GCC GTC AGA

    PUMA, forward: ATG CCT GCC TCA CCT TCA TC

    PUMA, reverse: TCA CAC GTC GCT CTC TCT AAA CC

    p21-1, forward: GCT GTG GCT CTG ATT GGC TTT

    p21-1, reverse: ACA GGC AGC CCA AGG ACA AA

    p21-2, forward: CAT CCC CAC AGC AGA GGA GAA

    p21-2, reverse: ACC CAG GCT TGG AGC AGC TA

    p21-3, forward: GAG TCC TGT TTG CTT CTG GGC A

    p21-3, reverse: CTG CAT TGG GGC TGC CTA TGT A

    PCNA, forward: CCA CCA TAA AGC TGG GGC TT

    PCNA, reverse: TCT CCC CGC CTC TTT GAC TC

  • ICC/IF image of ab1101 stained human HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1101, 5 µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) ab150113 used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1:200 dilution for 1h. DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, Hek293 and MCF7 cells.

  • Immunohistochemistry (Frozen sections) using ab1101 (1:100) in human brain tissue.

  • Overlay histogram showing HeLa cells stained with ab1101 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab1101, 1:50 dilution) for 30 min at 22ºC. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1:500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • All lanes : Anti-p53 antibody [DO-1] (ab1101) at 1/1000 dilution

    Lane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 2 : DU 145 (Human prostate carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Predicted band size : 43.6 kDa
    Observed band size : 50 kDa (why is the actual band size different from the predicted?)

    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 5% Milk before being incubated with ab1101 overnight at 4°C. Antibody binding was detected using ab175783 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

  • All lanes : Anti-p53 antibody [DO-1] (ab1101) at 2.5 µg/ml

    Lane 1 : MDA-MB-361 (Human breast adenocarcinoma cell line) Whole Cell Lysate Whole Cell Lysate
    Lane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 3 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Predicted band size : 43.6 kDa
    Observed band size : 53 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 55 kDa (possible post-translational modification).


  • Predicted band size : 43.6 kDa
    Observed band size : 53 kDa (why is the actual band size different from the predicted?)
    Lane 1 = Extract of Hek293T cells incubated with vehicle ? 20 ug. Lane 2 = Extract of Hek293T cells incubated with etoposide ? 20 ug. Lane 3 = Extract of Hek293T incubated with etoposide ? 7.5 ug. Lane 4 = Lambda phosphatase (400 times-diluted)-treated extract of Hek293T incubated with etoposide -7.5 ug. Lane 5 = Lambda phosphatase (100 times-diluted)-treated extract of Hek293T incubated with etoposide -7.5 ug. Lane 6 = Lambda phosphatase (25 times-diluted)-treated extract of Hek293T incubated with etoposide -7.5 ug. Lane 7 = Extract of Mcf7 cells incubated with vehicle ? 20 ug. Lane 8 = Extract of Mcf7 cells incubated 6 hours with camptothecin ? 20 ug. Lane 9 = Extract of Mcf7 cells incubated 16 hours with camptothecin ? 20 ug. Lane 10 = Extract of Mcf7 cells incubated 24 hours with camptothecin ? 20 ug. SDS PAGE performed under reducing conditions (100mM DTT ? Sample heated at 50?C).
    Primary : Lanes 1-10: Anti p53 antibody (ab1101) at 0.4 ug/mL Secondary : Lanes 1-10: Goat anti mouse IgG(H&L)-HRP at 1:10000. Developed: ECL with 30 sec exposure. Blocking: in 5% Milk in PBS for 3 hours at RT. Primary antibody: in 5% BSA in PBS overnight at 4 C. Secondary antibody: in 5% Milk in PBS for 2 hour at RT. Predicted band size : 53kDa. Observed band size : 53kDa
  • All lanes : Anti-p53 antibody [DO-1] (ab1101) at 1/1000 dilution

    Lane 1 : UV treated Human Lymphocytes
    Lane 2 : UV treated Human Lymphocytes

    Lysates/proteins at 30 µg per lane.

    Secondary
    Goat Polyclonal to Rabbit IgG at 1/2000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 43.6 kDa
    Observed band size : 53 kDa (why is the actual band size different from the predicted?)


    Exposure time : 10 seconds

    This image is courtesy of an anonymous abreview.

    See Abreview

Anti-p53 antibody [DO-1] (ab1101) 使用論文

This product has been referenced in:
  • Yang K  et al. Effect of PLCe gene silencing on inhibiting the cancerous transformation of ulcerative colitis. Exp Ther Med 12:422-426 (2016). WB ; Mouse . Read more (PubMed: 27347072) »
  • Hong X  et al. SOX9 is targeted for proteasomal degradation by the E3 ligase FBW7 in response to DNA damage. Nucleic Acids Res N/A:N/A (2016). Read more (PubMed: 27566146) »

See all 14 Publications for this product

Product Wall

Application Immunoprecipitation
Immuno-precipitation step Other - Protein G dynabeads
Sample Human Cell lysate - whole cell (HCT116 - with His tagged p53)
Specification HCT116 - with His tagged p53
Total protein in input 200 µg
Username

Neil Grayneil

Verified customer

投稿 Dec 23 2013

Thank you for contacting us.

This antibody was made against recombinant full-length human p53 corresponding to the sequence of native human p53. Positive controls can be whole cell lysates of 293T cells or A431 cells. The specific epitope re...

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Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Tissue lysate - whole (hepatocytes)
Specification hepatocytes
Treatment 100 U/ml IL-1
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

投稿 Jun 19 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (4-12%)
Sample Human Cell lysate - whole cell (lymphocytes)
Specification lymphocytes
Treatment UV treated
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Apr 07 2014

Application Western blot
Loading amount 100 µg
Gel Running Conditions Reduced Denaturing (Gradient Gel)
Sample Human Cell lysate - whole cell (HCT116)
Specification HCT116
Blocking step Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Abcam user community

Verified customer

投稿 Dec 30 2013

Application Western blot
Loading amount 20 µg
Gel Running Conditions Non-reduced Denaturing (10)
Sample Human Cell lysate - whole cell (HeLa)
Specification HeLa
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 Nov 13 2013

Thank you for contacting us. Unfortunately, as we carry over 70,000 products, it simply isn't feasible for us to keep small sample sizes of our products.

However, we are happy to reassure our customers that all of our products are covered by...

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Thank you for contacting us.

The number of tests totally depends on the dilution or concentration range customer uses. However at the recommended dilution, the antibody can be used for 100 ICC/IF or 100 IHC-P or 100 WB tests.

I hope t...

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I am sorry we are unable to resolve your inquiry at Abcam. Please contact our distributor Kimera with all the details. They will be able to resolve the case for you.

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ab1101 has been developed against the recombinant protein and has alsobeen tested with the endogenous protein (see http://www.abcam.com/p53-antibody-ab1101.html#p53-Primary-antibodies-ab1101-2.jpg). Therefore this antibody recognises both recombin...

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