Anti-HAUSP / USP7 antibody (ab101648)
Key features and details
- Rabbit polyclonal to HAUSP / USP7
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-HAUSP / USP7 antibody
See all HAUSP / USP7 primary antibodies -
Description
Rabbit polyclonal to HAUSP / USP7 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB MouseHuman
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Immunogen
Synthetic peptide corresponding to Human HAUSP/ USP7 aa 1-100 conjugated to keyhole limpet haemocyanin.
(Peptide available asab115932) -
Positive control
- This antibody gave a positive signal in the following whole cell lysates: HAP1 (WT); HeLa; HEK293; A431; MCF7; Ramos; PC3; as well as HeLa nuclear extract, Mouse Testis tissue lysate and Rat Testis tissue lysate. ICC/IF: HepG2 cells.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab101648 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanWB MouseHumanAll applications HorsePigMacaque monkeyApplication Abreviews Notes WB Use a concentration of 1 µg/ml. Detects a band of approximately 140 kDa (predicted molecular weight: 128 kDa).ICC/IF Use a concentration of 5 µg/ml.Notes WB
Use a concentration of 1 µg/ml. Detects a band of approximately 140 kDa (predicted molecular weight: 128 kDa).ICC/IF
Use a concentration of 5 µg/ml.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 (ab101648)
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 - ab101648 observed at 115 kDa. Red - loading control, ab8245, observed at 37 kDa.ab101648 was shown to recognize HAUSP/USP7 when HAUSP/USP7 knockout samples were used, along with additional cross-reactive bands. Wild-type and HAUSP/USP7 knockout samples were subjected to SDS-PAGE. ab101648 and ab8245 (loading control to GAPDH) were diluted 1 μg/mL and 1/2000 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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Western blot - Anti-HAUSP / USP7 antibody (ab101648)All lanes : Anti-HAUSP / USP7 antibody (ab101648) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 4 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 6 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate
Lane 7 : PC-3 whole cell lysate (ab3954)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 128 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?
Additional bands at: 68 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 2 minutes -
Immunocytochemistry/ Immunofluorescence - Anti-HAUSP / USP7 antibody (ab101648)ICC/IF image of ab101648 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab101648 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Western blot - Anti-HAUSP / USP7 antibody (ab101648)All lanes : Anti-HAUSP / USP7 antibody (ab101648) at 1 µg/ml
Lane 1 : Testis (Mouse) Tissue Lysate
Lane 2 : Testis (Rat) Tissue Lysate - normal tissue (ab29388)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 128 kDa
Observed band size: 128 kDa
Additional bands at: 45 kDa, 60 kDa, 90 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 2 minutes
Protocols
Datasheets and documents
References (1)
ab101648 has been referenced in 1 publication.
- Sin TK et al. Resveratrol protects against doxorubicin-induced cardiotoxicity in aged hearts through the SIRT1-USP7 axis. J Physiol N/A:N/A (2015). WB ; Mouse . PubMed: 25639460
Images
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Western blot - Anti-HAUSP / USP7 antibody (ab101648)
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 - ab101648 observed at 115 kDa. Red - loading control, ab8245, observed at 37 kDa.ab101648 was shown to recognize HAUSP/USP7 when HAUSP/USP7 knockout samples were used, along with additional cross-reactive bands. Wild-type and HAUSP/USP7 knockout samples were subjected to SDS-PAGE. ab101648 and ab8245 (loading control to GAPDH) were diluted 1 μg/mL and 1/2000 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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Western blot - Anti-HAUSP / USP7 antibody (ab101648)All lanes : Anti-HAUSP / USP7 antibody (ab101648) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 4 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 6 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate
Lane 7 : PC-3 whole cell lysate (ab3954)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 128 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?
Additional bands at: 68 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 2 minutes
-
Immunocytochemistry/ Immunofluorescence - Anti-HAUSP / USP7 antibody (ab101648)
ICC/IF image of ab101648 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab101648 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Western blot - Anti-HAUSP / USP7 antibody (ab101648)All lanes : Anti-HAUSP / USP7 antibody (ab101648) at 1 µg/ml
Lane 1 : Testis (Mouse) Tissue Lysate
Lane 2 : Testis (Rat) Tissue Lysate - normal tissue (ab29388)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 128 kDa
Observed band size: 128 kDa
Additional bands at: 45 kDa, 60 kDa, 90 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 2 minutes
