製品の概要

  • 製品名

    Anti-p73 antibody [EPR18409(T)(MIX)]
    p73 一次抗体 製品一覧
  • 製品の詳細

    Rabbit monoclonal [EPR18409(T)(MIX)] to p73
  • 由来種

    Rabbit
  • 特異性

    The immunogen used for this product shares 76% homology with p63. Cross-reactivity with this protein has not been confirmed experimentally.
  • アプリケーション

    適用あり: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • 種交差性

    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide within Human p73 aa 450-550. The exact sequence is proprietary.
    Database link: O15350

  • ポジティブ・コントロール

    • WB: Recombinant fragment of human p73 protein; HeLa, Jurkat, K562, WEHI-3, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Human fetal brain and fetal kidney lysates; Mouse, brain, heart, kidney and spleen lysates; Rat brain and spleen lysates. IHC-P: Human testis, glioma and gastric carcinoma tissues. ICC/IF: MCF7 and HeLa cells. Flow Cyt: Jurkat and K562 cells. IP: HeLa and MCF7 whole cell lysates.
  • 特記事項

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab189896 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
WB 1/5000. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/1000.
Flow Cyt 1/300.
IP 1/80.

ターゲット情報

  • 機能

    Participates in the apoptotic response to DNA damage. Isoforms containing the transactivation domain are pro-apoptotic, isoforms lacking the domain are anti-apoptotic and block the function of p53 and transactivating p73 isoforms. May be a tumor suppressor protein.
  • 組織特異性

    Expressed in striatal neurons of patients with Huntington disease (at protein level). Brain, kidney, placenta, colon, heart, liver, spleen, skeletal muscle, prostate, thymus and pancreas. Highly expressed in fetal tissue.
  • 配列類似性

    Belongs to the p53 family.
    Contains 1 SAM (sterile alpha motif) domain.
  • ドメイン

    Possesses an acidic transactivation domain, a central DNA binding domain and a C-terminal oligomerization domain that binds to the ABL tyrosine kinase SH3 domain.
    The WW-binding motif mediates interaction with WWOX.
  • 翻訳後修飾

    Isoform alpha (but not isoform beta) is sumoylated on Lys-627, which potentiates proteasomal degradation but does not affect transcriptional activity.
    Higher levels of phosphorylation seen in the brain from patients with Huntington disease.
    Ubiquitinated; leading to its degradation by the proteasome.
  • 細胞内局在

    Nucleus. Accumulates in the nucleus in response to DNA damage.
  • Information by UniProt
  • 参照データベース

  • 別名

    • p53 like transcription factor antibody
    • p53 related protein antibody
    • p53-like transcription factor antibody
    • p53-related protein antibody
    • p73 antibody
    • P73_HUMAN antibody
    • TP73 antibody
    • Tumor protein p73 antibody
    see all

画像

  • Anti-p73 antibody [EPR18409(T)(MIX)] (ab189896) at 1/5000 dilution + Recombinant fragment of human p73 protein at 0.005 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 70 kDa
    Observed band size: 28 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Recombinant fragment of human p73 protein contains aa380-636 with His-Tag®.

  • All lanes : Anti-p73 antibody [EPR18409(T)(MIX)] (ab189896) at 1/5000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 3 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
    Lane 4 : WEHI-3 (Mouse leukemia cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 70 kDa
    Observed band size: 70 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-p73 antibody [EPR18409(T)(MIX)] (ab189896) at 1/5000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/50000 dilution

    Predicted band size: 70 kDa
    Observed band size: 70 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-p73 antibody [EPR18409(T)(MIX)] (ab189896) at 1/5000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Mouse spleen lysate
    Lane 5 : Rat brain lysate
    Lane 6 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 70 kDa
    Observed band size: 70 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-p73 antibody [EPR18409(T)(MIX)] (ab189896) at 1/5000 dilution

    Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 70 kDa
    Observed band size: 70 kDa


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling p73 alpha+beta with ab189896 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Negative staining on Human cerebral cortex.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling p73 alpha+beta with ab189896 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic and weak nucleus staining on Human spermatogonial cells is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Human glioma tissue labeling p73 alpha+beta with ab189896 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nucleus and cytoplasmic staining on Human glioma is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Human gastric carcinoma tissue labeling p73 alpha+beta with ab189896 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nucleus and cytoplasmic staining on Human gastric carcinoma is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling p73 alpha+beta with ab189896 at 1/1000 dilution, followed by Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear and cytoplasm staining on MCF7 cell line. 

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody-Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab189896 at 1/1000 dilution followed by ab150120 at 1/1000 dilution.

    -ve control 2: ab7291at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling p73 alpha+beta with ab189896 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic and weakly nuclear staining on HeLa cell line. 

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (AlexaFluor®594) preadsorbed (ab150120) at  1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab189896 at 1/1000 dilution followed by ab150120 at 1/1000 dilution.

    -ve control 2: ab7291 at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling p73 alpha+beta with ab189896 at 1/300 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed K562 (Human chronic myelogenous leukemia cell line from bone marrow) cells labeling p73 alpha+beta with ab189896 at 1/300 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

  • p73 alpha+beta was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab189896 at 1/80 dilution.

    Western blot was performed from the immunoprecipitate using ab189896 at 1/1000 dilution.

    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500.

    Lane 1: HeLa whole cell lysate 10ug (Input).

    Lane 2: ab189896 IP in HeLa whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab189896 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

  • p73 alpha+beta was immunoprecipitated from 1mg of MCF7 (Human breast adenocarcinoma cell line) whole cell lysate with ab189896 at 1/80 dilution.

    Western blot was performed from the immunoprecipitate using ab189896 at 1/1000 dilution.

    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500.

    Lane 1: MCF7 whole cell lysate 10ug (Input).

    Lane 2: ab189896 IP in MCF7 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab189896 in MCF7 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

参考文献

This product has been referenced in:

  • Iannolo G  et al. Zika virus infection induces MiR34c expression in glioblastoma stem cells: new perspectives for brain tumor treatments. Cell Death Dis 10:263 (2019). Read more (PubMed: 30890698) »
  • El Husseini N & Hales BF The Roles of P53 and Its Family Proteins, P63 and P73, in the DNA Damage Stress Response in Organogenesis-Stage Mouse Embryos. Toxicol Sci 162:439-449 (2018). Read more (PubMed: 29228353) »
See all 2 Publications for this product

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1-3 of 3 Abreviews

Application
Western blot
Sample
Human Cell lysate - whole cell (MCF7 (breast cancer), HepG2 (liver cancer), 293T ()
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Loading amount
12 µg
Specification
MCF7 (breast cancer), HepG2 (liver cancer), 293T (
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

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投稿 Nov 01 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (MDA-MB-231)
Permeabilization
Yes - Tween-20
Specification
MDA-MB-231
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Acetone

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投稿 Jan 09 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (paraffin-embedded human lung carcinoma)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 6.0 Citric Acid
Permeabilization
Yes - Tween-20
Specification
paraffin-embedded human lung carcinoma
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

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投稿 Jan 09 2018

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