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Cell Biology Cell Cycle Cell Cycle Inhibitors p53
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)

  • Datasheet
Reviews (2) Submit a question References (1)

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Western blot - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
  • Flow Cytometry - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
  • Western blot - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
  • Western blot - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
  • Immunocytochemistry/ Immunofluorescence - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
  • Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4253] to HAUSP / USP7
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

You may also be interested in

Protein
Product image
Recombinant human HAUSP / USP7 protein (ab198464)
Knockout
Product image
Human USP7 (HAUSP) knockout HEK-293T cell line (ab266535)
Primary
Product image
Anti-p95/NBS1 antibody [Y112] (ab32074)

View more associated products

Overview

  • Product name

    Anti-HAUSP / USP7 antibody [EPR4253]
    See all HAUSP / USP7 primary antibodies
  • Description

    Rabbit monoclonal [EPR4253] to HAUSP / USP7
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

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

  • Positive control

    • WB: HEK293T, Ramos, HeLa, A431, MCF7, C6 and RAW 264.7 cell lysates. IHC-P: Human colon tissue. IF: HeLa cells. Flow Cyt: HeLa cells.
  • 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR4253
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • p53
    • Microbiology
    • Interspecies Interaction
    • Host Virus Interaction
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Deubiquitination
    • Cancer
    • Cell cycle
    • Cell cycle inhibitors
    • p53 pathway
    • Epigenetics and Nuclear Signaling
    • Ubiquitin & Ubiquitin Like Modifiers
    • Deubiquitination

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-HAUSP / USP7 antibody [EPR4253] (ab214647)
    • Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human USP7 (HAUSP) knockout HEK-293T cell line (ab266535)
  • KO cell lysates

    • Human USP7 (HAUSP) knockout HEK-293T cell lysate (ab257284)
  • Positive Controls

    • HeLa nuclear extract lysate (ab14655)
  • Recombinant Protein

    • Recombinant human HAUSP / USP7 protein (ab198464)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab108931 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
WB
Human
Application Abreviews Notes
WB (1)
1/1000 - 1/10000. Predicted molecular weight: 128 kDa.
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Antigen retrieval is recommended.

ICC/IF (1)
1/100 - 1/250.
Flow Cyt
Use at an assay dependent concentration.
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 128 kDa.
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Antigen retrieval is recommended.

ICC/IF
1/100 - 1/250.
Flow Cyt
Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function

      Hydrolase that deubiquitinates target proteins such as FOXO4, p53/TP53, MDM2, ERCC6, DNMT1, UHRF1, PTEN and DAXX (PubMed:11923872, PubMed:15053880, PubMed:16964248, PubMed:18716620, PubMed:25283148). Together with DAXX, prevents MDM2 self-ubiquitination and enhances the E3 ligase activity of MDM2 towards p53/TP53, thereby promoting p53/TP53 ubiquitination and proteasomal degradation. Deubiquitinates p53/TP53, preventing degradation of p53/TP53, and enhances p53/TP53-dependent transcription regulation, cell growth repression and apoptosis (PubMed:25283148). Deubiquitinates p53/TP53 and MDM2 and strongly stabilizes p53/TP53 even in the presence of excess MDM2, and also induces p53/TP53-dependent cell growth repression and apoptosis. Deubiquitination of FOXO4 in presence of hydrogen peroxide is not dependent on p53/TP53 and inhibits FOXO4-induced transcriptional activity. In association with DAXX, is involved in the deubiquitination and translocation of PTEN from the nucleus to the cytoplasm, both processes that are counteracted by PML. Involved in cell proliferation during early embryonic development. Involved in transcription-coupled nucleotide excision repair (TC-NER) in response to UV damage: recruited to DNA damage sites following interaction with KIAA1530/UVSSA and promotes deubiquitination of ERCC6, preventing UV-induced degradation of ERCC6. Contributes to the overall stabilization and trans-activation capability of the herpesvirus 1 trans-acting transcriptional protein ICP0/VMW110 during HSV-1 infection. Involved in maintenance of DNA methylation via its interaction with UHRF1 and DNMT1: acts by mediating deubiquitination of UHRF1 and DNMT1, preventing their degradation and promoting DNA methylation by DNMT1 (PubMed:21745816). Exhibits a preference towards 'Lys-48'-linked ubiquitin chains. Increases regulatory T-cells (Treg) suppressive capacity by deubiquitinating and stabilizing the transcription factor FOXP3 which is crucial for Treg cell function (PubMed:23973222).
    • Tissue specificity

      Widely expressed. Overexpressed in prostate cancer.
    • Sequence similarities

      Belongs to the peptidase C19 family.
      Contains 1 MATH domain.
      Contains 1 USP domain.
    • Domain

      The C-terminus plays a role in its oligomerization.
    • Post-translational
      modifications

      Isoform 1: Phosphorylated. Isoform 1 is phosphorylated at positions Ser-18 and Ser-963. Isoform 2: Not phosphorylated.
      Isoform 1: Polyneddylated. Isoform 2: Not Polyneddylated.
      Isoform 1 and isoform 2: Not sumoylated.
      Isoform 1 and isoform 2: Polyubiquitinated by herpesvirus 1 trans-acting transcriptional protein ICP0/VMW110; leading to its subsequent proteasomal degradation. Isoform 1: Ubiquitinated at Lys-869.
    • Cellular localization

      Nucleus. Cytoplasm. Nucleus, PML body. Present in a minority of ND10 nuclear bodies. Association with ICP0/VMW110 at early times of infection leads to an increased proportion of USP7-containing ND10. Colocalizes with ATXN1 in the nucleus. Colocalized with DAXX in speckled structures. Colocalized with PML and PTEN in promyelocytic leukemia protein (PML) nuclear bodies.
    • Target information above from: UniProt accession Q93009 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 7874 Human
      • Entrez Gene: 252870 Mouse
      • Entrez Gene: 360471 Rat
      • Omim: 602519 Human
      • SwissProt: Q93009 Human
      • SwissProt: Q6A4J8 Mouse
      • SwissProt: Q4VSI4 Rat
      • Unigene: 386939 Human
      • Unigene: 295330 Mouse
      • Unigene: 72721 Rat
      see all
    • Alternative names

      • Deubiquitinating enzyme 7 antibody
      • HAUSP antibody
      • Herpes virus associated ubiquitin specific protease antibody
      • Herpesvirus-associated ubiquitin-specific protease antibody
      • TEF 1 antibody
      • tef-1 antibody
      • TEF1 antibody
      • Ubiquitin carboxyl terminal hydrolase 7 antibody
      • Ubiquitin carboxyl-terminal hydrolase 7 antibody
      • Ubiquitin specific peptidase 7 (herpes virus associated) antibody
      • Ubiquitin specific peptidase 7 antibody
      • Ubiquitin specific peptidase 7 herpes virus associated antibody
      • Ubiquitin specific processing protease 7 antibody
      • Ubiquitin specific protease 7 (herpes virus associated) antibody
      • Ubiquitin specific protease 7 antibody
      • Ubiquitin specific protease 7 herpes virus associated antibody
      • Ubiquitin thioesterase 7 antibody
      • Ubiquitin thiolesterase 7 antibody
      • Ubiquitin-specific-processing protease 7 antibody
      • UBP 7 antibody
      • UBP-7 antibody
      • UBP7 antibody
      • UBP7_HUMAN antibody
      • USP 7 antibody
      • usp-7 antibody
      • Usp7 antibody
      • VMW110-ASSOCIATED PROTEIN, 135-KD antibody
      see all

    Images

    • Western blot - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
      Western blot - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: HAUSP/USP7 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: MCF7 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab108931 observed at 115 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab108931 was shown to specifically react with HAUSP/USP7 when HAUSP/USP7 knockout samples were used. Wild-type and HAUSP/USP7 knockout samples were subjected to SDS-PAGE. ab108931 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    • Flow Cytometry - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
      Flow Cytometry - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)

      Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling HAUSP / USP7  with unpurified ab108931 at 1:30 dilution(10ug/ml) (red). Cells were fixed with 80% methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor® 488)(ab150077)(1:2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black)(ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)

      ab108931, at 1/50, staining HAUSP / USP7 in Human colon tissue by Immunohistochemistry, Paraffin-embedded tissue.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Western blot - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
      Western blot - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
      All lanes : Anti-HAUSP / USP7 antibody [EPR4253] (ab108931) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : USP7 knockout HEK-293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 128 kDa
      Observed band size: 128 kDa



      Lanes 1- 2: Merged signal (red and green). Green - ab108931 observed at 128 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab108931 was shown to react with HAUSP / USP7 in wild-type HEK-293T cells in western blot. The band observed in knockout cell line ab266535 (knockout cell lysate ab257284) lane below 128kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HEK-293T and USP7 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab108931 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
      Western blot - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
      All lanes : Anti-HAUSP / USP7 antibody [EPR4253] (ab108931) at 1/1000 dilution

      Lane 1 : Ramos cell lysate
      Lane 2 : HeLa cell lysate
      Lane 3 : A431 cell lysate
      Lane 4 : MCF7 cell lysate
      Lane 5 : C6 cell lysate
      Lane 6 : RAW 264.7 cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 128 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
      Immunocytochemistry/ Immunofluorescence - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
      ab108931, at 1/100, staining HAUSP / USP7 in HeLa cells by Immunofluorescence.
    • Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
      Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
  • References (1)

    Publishing research using ab108931? Please let us know so that we can cite the reference in this datasheet.

    ab108931 has been referenced in 1 publication.

    • Hu T  et al. Targeting the overexpressed USP7 inhibits esophageal squamous cell carcinoma cell growth by inducing NOXA-mediated apoptosis. Mol Carcinog 58:42-54 (2019). PubMed: 30182448

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-2 of 2 Abreviews or Q&A

    Western blot abreview for Anti-HAUSP / USP7 antibody [EPR4253]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HAP1 cells)
    Gel Running Conditions
    Non-reduced Denaturing (4-12 % Bis-Tris gel)
    Loading amount
    50 µg
    Specification
    HAP1 cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 20 2016

    Immunocytochemistry/ Immunofluorescence abreview for Anti-HAUSP / USP7 antibody [EPR4253]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.05% Triton X100 in PBS
    Read More

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Feb 20 2013

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