Recombinant Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
![Western blot - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)](https://www.abcam.com/ps/products/108/ab108931/Images/ab108931-256361-anti-hausp-usp7-antibody-epr4253-western-blot.jpg "Western blot - 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: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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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
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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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 (intra): HeLa cells.
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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
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
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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
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR4253 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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.
| Application | Abreviews | Notes |
|---|---|---|
| Flow Cyt (Intra) |
Use at an assay dependent concentration.
|
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| 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. |
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| ICC/IF | (1) |
1/100 - 1/250.
|
| Notes |
|---|
|
Flow Cyt (Intra)
Use at an assay dependent concentration. |
|
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. |
Target
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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
modificationsIsoform 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
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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
see all -
Alternative names
- Deubiquitinating enzyme 7 antibody
- HAUSP antibody
- Herpes virus associated ubiquitin specific protease antibody
see all
Images
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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.
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![Flow Cytometry (Intracellular) - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)](https://www.abcam.com/ps/products/108/ab108931/Images/ab108931-276878-anti-hausp-usp7-antibody-epr4253-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)](https://www.abcam.com/ps/products/108/ab108931/Images/ab108931-121425-anti-hausp-usp7-antibody-epr4253-immunohistochemistry.jpg)
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.
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![Western blot - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)](https://www.abcam.com/ps/products/108/ab108931/Images/ab108931-409572-anti-hausp-usp7-antibody-epr4253-western-blot-edited-hek293t.jpg)
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 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 kDaLanes 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.
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![Western blot - Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)](https://www.abcam.com/ps/products/108/ab108931/Images/ab108931-121426-anti-hausp-usp7-antibody-epr4253-western-blot.gif)
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)](https://www.abcam.com/ps/products/108/ab108931/Images/ab108931-121424-anti-hausp-usp7-antibody-epr4253-immunofluorescence.jpg)
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)](https://www.abcam.com/ps/products/108/ab108931/Images/ab108931-7-benefits-of-recombinant-antibodies.png)
Anti-HAUSP / USP7 antibody [EPR4253] (ab108931)
Protocols
Datasheets and documents
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Datasheet download
References (3)
ab108931 has been referenced in 3 publications.
- Liu J et al. Arginine methylation-dependent LSD1 stability promotes invasion and metastasis of breast cancer. EMBO Rep 21:e48597 (2020). PubMed: 31833203
- 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
- Xia X et al. Deubiquitination and stabilization of estrogen receptor a by ubiquitin-specific protease 7 promotes breast tumorigenesis. Cancer Lett 465:118-128 (2019). PubMed: 31518603
