Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)

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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 - BSA and Azide free
  • Suitable for: IHC-P, ICC/IF, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free
    See all HAUSP / USP7 primary antibodies

  • Description

    Rabbit monoclonal [EPR4253] to HAUSP / USP7 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, ICC/IF, Flow Cyt, WBmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human HAUSP/ USP7 aa 100-200. The exact sequence is proprietary.

  • Positive control

    • WB: HEK-29T cell lysate. Flow Cyt: HeLa cells. IHC-P: Human colon tissue. ICC: HeLa cells.

  • General notes

    Ab239936 is the carrier-free version of ab108931. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab239936 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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

Applications

Our Abpromise guarantee covers the use of ab239936 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
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Antigen retrieval is recommended.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 128 kDa.
  • 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.
    • Information by UniProt

    • Database links

      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] - BSA and Azide free (ab239936)
      Western blot - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)
      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



      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108931).

      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.

    • Flow Cytometry - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)
      Flow Cytometry - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108931).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108931).

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

    • Immunocytochemistry/ Immunofluorescence - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)
      Immunocytochemistry/ Immunofluorescence - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)
      ab108931, at 1/100, staining HAUSP / USP7 in HeLa cells by Immunofluorescence.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108931).

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab239936? Please let us know so that we can cite the reference in this datasheet.

    ab239936 has not yet been referenced specifically in any publications.

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