Validated using a knockout cell line
Recombinant
RabMAb

Anti-USP11 antibody [EPR4346] (ab109232)

Overview

  • Product name
    Anti-USP11 antibody [EPR4346]
    See all USP11 primary antibodies
  • Description
    Rabbit monoclonal [EPR4346] to USP11
  • Tested applications
    Suitable for: WB, IP, Flow Cytmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human USP11 aa 850-950. The exact sequence is proprietary.
    Database link: P51784

  • Positive control
    • WB: 293T, Jurkat, Human testis, Human fetal kidney and LnCaP cell lysates
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109232 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
WB 1/1000 - 1/10000. Detects a band of approximately 110 kDa (predicted molecular weight: 110 kDa).

 

IP 1/10 - 1/300.

 

Flow Cyt 1/500 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Function
      Protease that can remove conjugated ubiquitin from target proteins and polyubiquitin chains. Inhibits the degradation of target proteins by the proteasome. Plays a role in the regulation of pathways leading to NF-kappa-B activation. Plays a role in the regulation of DNA repair after double-stranded DNA breaks.
    • Sequence similarities
      Belongs to the peptidase C19 family.
      Contains 1 DUSP domain.
    • Cellular localization
      Nucleus. Cytoplasm. Predominantly nuclear. Associates with chromatin.
    • Information by UniProt
    • Database links
    • Alternative names
      • Deubiquitinating enzyme 11 antibody
      • Ubiquitin carboxyl-terminal hydrolase 11 antibody
      • Ubiquitin carboxyl-terminal hydrolase X linked antibody
      • ubiquitin specific peptidase 11 antibody
      • Ubiquitin specific protease 11 antibody
      • Ubiquitin thiolesterase 11 antibody
      • Ubiquitin-specific-processing protease 11 antibody
      • UBP11_HUMAN antibody
      • UHX1 antibody
      • USP11 antibody
      see all

    Images



    • Predicted band size : 110 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: USP11 knockout HAP1 cell lysate (20 µg)
      Lane 3: LNCaP cell lysate (20 µg)
      Lane 4: HeLa cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab109232 observed at 110 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab109232 was shown to specifically react with USP11 when USP11 knockout samples were used. Wild-type and USP11 knockout samples were subjected to SDS-PAGE. ab109232 and ab8245 (loading control to GAPDH) were diluted 1/5000 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.

    • Overlay histogram showing Jurkat cells fixed in 4% PFA and stained with purified ab109232 at a dilution of 1 in 950 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
    • ab109232 (purified) at 1/300 immunoprecipitating USP11 in 10 μg HEK293 (Lanes 1 and 2, observed at 110 kDa). Lane 3 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
    • All lanes : Anti-USP11 antibody [EPR4346] (ab109232) at 1/5000 dilution (purified)

      Lane 1 : mouse brain lysate
      Lane 2 : rat brain lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      HRP goat anti-rabbit IgG (H+L) at 1000 µg

      Predicted band size : 110 kDa
      Observed band size : 110 kDa

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Flow cytometry analysis of 2% paraformaldehyde fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling USP11 with unpurified ab109232 at 1/570 dilution (red line). Secondary antibody used is a goat anti rabbit IgG (FITC) at 1/150 dilution. The isotype control is rabbit monoclonal IgG (black line). The unlabeled control is cells without incubation with primary and secondary antibodies (blue line).

    • ab109232 (purified) at 1/300 immunoprecipitating USP11 in 10 μg Jurkat (Lanes 1 and 2, observed at 110 kDa). Lane 3 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
    • All lanes : Anti-USP11 antibody [EPR4346] (ab109232) at 1/5000 dilution (purified)

      Lane 1 : HEK293 lysate at 20 µg
      Lane 2 : human fetal kidney lysate

      Secondary
      HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 110 kDa
      Observed band size : 110 kDa

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • All lanes : Anti-USP11 antibody [EPR4346] (ab109232) at 1/1000 dilution (purified)

      Lane 1 : Jurkat lysate
      Lane 2 : human testis lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 110 kDa
      Observed band size : 110 kDa

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • All lanes : Anti-USP11 antibody [EPR4346] (ab109232) at 1/1000 dilution (unpurified)

      Lane 1 : 293T cell lysate
      Lane 2 : Jurkat cell lysate
      Lane 3 : Human testis cell lysate
      Lane 4 : Huma fetal kidney cell lysate
      Lane 5 : LnCaP cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size : 110 kDa
      Observed band size : 110 kDa

    References

    This product has been referenced in:
    • Orthwein A  et al. A mechanism for the suppression of homologous recombination in G1 cells. Nature 528:422-6 (2015). IP ; Human . Read more (PubMed: 26649820) »
    • Laurette P  et al. Transcription factor MITF and remodeller BRG1 define chromatin organisation at regulatory elements in melanoma cells. Elife 4:N/A (2015). WB ; Human . Read more (PubMed: 25803486) »

    See all 2 Publications for this product

    Customer reviews and Q&As

    Application
    Flow Cytometry
    Fixation
    ebioscience Fixation/Permeablization kit
    Permeabilization
    Yes - ebioscience Fixation/Permeablization kit
    Sample
    Mouse Cell (Lymphocytes from peripheral lymph nodes and spleen)
    Specification
    Lymphocytes from peripheral lymph nodes and spleen
    Gating Strategy
    Lymphocytes were selected using FSC and SSC followed by gating on viable cells.
    Preparation
    Cell harvesting/tissue preparation method: Extracting spleen and lymph nodes during necropsy, followed by mashing through 70 micron filter
    Sample buffer: Sample collected in PBS
    Username

    Roman Istomine

    Verified customer

    Submitted Oct 22 2014

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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