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    hausp--usp7-antibody-epr4253-bsa-and-azide-free-ab239936.pdf

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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] - BSA and Azide free (ab239936)

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Western blot - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)
  • Flow Cytometry (Intracellular) - 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)
  • Western blot - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)
  • Immunocytochemistry/ Immunofluorescence - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)
  • Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)

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, Flow Cyt (Intra), ICC/IF, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Protein
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Recombinant human HAUSP / USP7 protein (ab198464)

View more associated products

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, Flow Cyt (Intra), ICC/IF, WBmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control

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

    ab239936 is the carrier-free version of ab108931.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    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.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • 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

    • Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
    • Alexa Fluor® 488 Anti-HAUSP / USP7 antibody [EPR4253] (ab214647)
    • PE Anti-HAUSP / USP7 antibody [EPR4253] (ab303172)
    • APC Anti-HAUSP / USP7 antibody [EPR4253] (ab303173)
    • HRP Anti-HAUSP / USP7 antibody [EPR4253] (ab303174)
  • Compatible Secondaries

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

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • 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)
  • Recombinant Protein

    • Recombinant human HAUSP / USP7 protein (ab198464)

Applications

The Abpromise guarantee

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.

Flow Cyt (Intra)
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 128 kDa.
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.

Flow Cyt (Intra)
Use at an assay dependent concentration.
ICC/IF
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.
  • 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] - BSA and Azide free (ab239936)
    Western blot - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)

    This data was developed using the same antibody clone in a different buffer formulation (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 (Intracellular) - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)
    Flow Cytometry (Intracellular) - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (ab239936)

    Intracellular 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.

  • 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 CRISPR/Cas9 edited 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 CRISPR/Cas9 edited cell line ab266535 (CRISPR/Cas9 edited 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 CRISPR/Cas9 edited 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.

  • 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).

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

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

Datasheets and documents

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

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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