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USP7 Recombinant Rabbit Monoclonal Antibody [JB80-36]

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Specification

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Catalog# ET7107-53

USP7 Recombinant Rabbit Monoclonal Antibody [JB80-36]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

USP7 Recombinant Rabbit Monoclonal Antibody [JB80-36]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human USP7 aa 80-180 / 1,102.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 128 kDa

Positive Control

A375 cell lysate, NIH/3T3 cell lysate, 4T1 cell lysate, C6 cell lysate, Mouse colon tissue lysate, Mouse testis tissue lysate, Rat testis tissue lysate, 293T cell lysate, HeLa cell lysate, human tonsils tissue, human testis tissue, rat brain tissue.

Conjugation

unconjugated

Clone Number

JB80-36

RRID

AB_3070762

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:1,000

  • IHC-P

  • 1:200

Target

Function

Ubiquitin-specific-processing protease 7 (USP7), also known as ubiquitin carboxyl-terminal hydrolase 7 or herpesvirus-associated ubiquitin-specific protease (HAUSP), is an enzyme that in humans is encoded by the USP7 gene. USP7 or HAUSP is a ubiquitin specific protease or a deubiquitylating enzyme that cleaves ubiquitin from its substrates.[9] Since ubiquitylation (polyubiquitination) is most commonly associated with the stability and degradation of cellular proteins, HAUSP activity generally stabilizes its substrate proteins. Another important role of HAUSP function involves the oncogenic stabilization of p53. Oncogenes such as Myc and E1A are thought to activate p53 through a p19 alternative reading frame (p19ARF, also called ARF)-dependent pathway, although some evidence suggests ARF is not essential in this process. A possibility is that HAUSP provides an alternative pathway for safeguarding the cell against oncogenic insults. USP7 can deubiquitinate histone H2B and this activity is associated with gene silencing in Drosophila. USP7 associates with a metabolic enzyme, GMP synthetase (GMPS) and this association stimulates USP7 deubiquitinase activity towards H2B. The USP7-GMPS complex is recruited to the polycomb (Pc) region in Drosophila and contributes to epigenetic silencing of homeotic genes.

Background References

1. Li M et al. Deubiquitination of p53 by HAUSP is an important pathway for p53 stabilization. Nature 416:648-653 (2002).

2. van der Horst A et al. FOXO4 transcriptional activity is regulated by monoubiquitination and USP7/HAUSP. Nat Cell Biol 8:1064-1073 (2006).

Sequence Similarity

Belongs to the peptidase C19 family.

Tissue Specificity

Expressed in neural progenitor cells (at protein level). Widely expressed. Overexpressed in prostate cancer.

Post-translational Modification

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.

Subcellular Location

Nucleus. Cytoplasm.

UNIPROT

SwissProt: Q93009 Human

SwissProt: Q6A4J8 Mouse

SwissProt: Q4VSI4 Rat

Synonyms

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

Expand

Images

  • <span style="font-weight: bold;">☑ Knockout (KO)</span><br /><br />All lanes: Western blot analysis of USP7 with anti-USP7 antibody [JB80-36] (<a href="/products/ET7107-53" style="font-weight: bold;text-decoration: underline;">ET7107-53</a>) at 1:1,000 dilution.<br />Lane 1: Wild-type A375 whole cell lysate (20 µg).<br />Lane 2: USP7 knockout A375 whole cell lysate (20 µg).<br /><br /><a href="/products/ET7107-53" style="font-weight: bold;text-decoration: underline;">ET7107-53</a> was shown to specifically react with USP7 in wild-type A375 cells. No band was observed when USP7 knockout sample was tested. Wild-type and USP7 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7107-53" style="font-weight: bold;text-decoration: underline;">ET7107-53</a>, 1/1,000) and Loading control antibody (Rabbit anti-HSP90, <a href="/products/ET1605-56" style="font-weight: bold;text-decoration: underline;">ET1605-56</a>, 1/10,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:200,000 dilution was used for 1 hour at room temperature.

    ☑ Knockout (KO)

    All lanes: Western blot analysis of USP7 with anti-USP7 antibody [JB80-36] (ET7107-53) at 1:1,000 dilution.
    Lane 1: Wild-type A375 whole cell lysate (20 µg).
    Lane 2: USP7 knockout A375 whole cell lysate (20 µg).

    ET7107-53 was shown to specifically react with USP7 in wild-type A375 cells. No band was observed when USP7 knockout sample was tested. Wild-type and USP7 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET7107-53, 1/1,000) and Loading control antibody (Rabbit anti-HSP90, ET1605-56, 1/10,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of USP7 on different lysates with Rabbit anti-USP7 antibody (<a href="/products/ET7107-53" style="font-weight: bold;text-decoration: underline;">ET7107-53</a>) at 1/1,000 dilution.<br /><br />Lane 1: NIH/3T3 cell lysate<br />Lane 2: 4T1 cell lysate<br />Lane 3: C6 cell lysate<br />Lane 4: Mouse colon tissue lysate<br />Lane 5: Mouse testis tissue lysate<br />Lane 6: Rat testis tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 128 kDa<br />Observed band size: 128 kDa<br /><br />Exposure time: 25 seconds; ECL: K1802;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7107-53" style="font-weight: bold;text-decoration: underline;">ET7107-53</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of USP7 on different lysates with Rabbit anti-USP7 antibody (ET7107-53) at 1/1,000 dilution.

    Lane 1: NIH/3T3 cell lysate
    Lane 2: 4T1 cell lysate
    Lane 3: C6 cell lysate
    Lane 4: Mouse colon tissue lysate
    Lane 5: Mouse testis tissue lysate
    Lane 6: Rat testis tissue lysate

    Lysates/proteins at 20 µg/Lane.

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

    Exposure time: 25 seconds; ECL: K1802;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7107-53) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of USP7 on different lysates with Rabbit anti-USP7 antibody (<a href="/products/ET7107-53" style="font-weight: bold;text-decoration: underline;">ET7107-53</a>) at 1/1,000 dilution.<br /><br />Lane 1: 293T cell lysate<br />Lane 2: HeLa cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 128 kDa<br />Observed band size: 128 kDa<br /><br />Exposure time: 2 minutes;<br /><br />6% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7107-53" style="font-weight: bold;text-decoration: underline;">ET7107-53</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of USP7 on different lysates with Rabbit anti-USP7 antibody (ET7107-53) at 1/1,000 dilution.

    Lane 1: 293T cell lysate
    Lane 2: HeLa cell lysate

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 2 minutes;

    6% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7107-53) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human tonsils tissue with Rabbit anti-USP7 antibody (<a href="/products/ET7107-53" style="font-weight: bold;text-decoration: underline;">ET7107-53</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7107-53" style="font-weight: bold;text-decoration: underline;">ET7107-53</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human tonsils tissue with Rabbit anti-USP7 antibody (ET7107-53) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-53) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-USP7 antibody (<a href="/products/ET7107-53" style="font-weight: bold;text-decoration: underline;">ET7107-53</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7107-53" style="font-weight: bold;text-decoration: underline;">ET7107-53</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-USP7 antibody (ET7107-53) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-53) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-USP7 antibody (<a href="/products/ET7107-53" style="font-weight: bold;text-decoration: underline;">ET7107-53</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7107-53" style="font-weight: bold;text-decoration: underline;">ET7107-53</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-USP7 antibody (ET7107-53) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-53) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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