Overview

  • Product name
    Anti-DUSP6 antibody [EPR129Y]
    See all DUSP6 primary antibodies
  • Description
    Rabbit monoclonal [EPR129Y] to DUSP6
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, IHC-FoFr, WB, IP, Flow Cytmore details
    Unsuitable for: ICC/IF
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human DUSP6 aa 350 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: Q16828
    (Peptide available as ab171765)

  • Positive control
    • WB: 3T3 cell lysate IHC: Human gastric carcinoma tissue.
  • 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

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR129Y
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab76310 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 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-FoFr Use at an assay dependent concentration.
WB 1/500 - 1/1000. Detects a band of approximately 42,44 kDa (predicted molecular weight: 42 kDa).Can be blocked with DUSP6 peptide (ab171765).
IP 1/40 - 1/50.
Flow Cyt 1/20 - 1/200.

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

  • Application notes
    Is unsuitable for ICC/IF.
  • Target

    • Function
      Inactivates MAP kinases. Has a specificity for the ERK family.
    • Sequence similarities
      Belongs to the protein-tyrosine phosphatase family. Non-receptor class dual specificity subfamily.
      Contains 1 rhodanese domain.
      Contains 1 tyrosine-protein phosphatase domain.
    • Cellular localization
      Cytoplasm.
    • Information by UniProt
    • Database links
    • Alternative names
      • Dual specificity phosphatase 6 antibody
      • Dual specificity phosphatase 6 isoform a antibody
      • Dual specificity protein phosphatase 6 antibody
      • Dual specificity protein phosphatase PYST1 antibody
      • DUS6_HUMAN antibody
      • DUSP 6 antibody
      • DUSP 6a antibody
      • Dusp6 antibody
      • DUSP6a antibody
      • HH19 antibody
      • MAP kinase phosphatase 3 antibody
      • Mitogen activated protein kinase phosphatase 3 antibody
      • Mitogen-activated protein kinase phosphatase 3 antibody
      • MKP 3 antibody
      • MKP-3 antibody
      • MKP3 antibody
      • PYST 1 antibody
      • PYST1 antibody
      • Serine/threonine specific protein phosphatase antibody
      see all

    Images

    • Immunohistochemical staining of paraffin embedded human pancreas with purified ab76310 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
    • Immunofluorescence staining of PC-12 cells with purified ab76310 at a working dilution of 1/200, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab76310 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

    • Immunoblot analysis of indicated proteins in splenocytes after CD4+ sort, liver homogenate or purified CD4+ T cells from WT, DUSP6+/- or DUSP6-/- mice to confirm knockout or heterozygosity for DUSP6 at protein level.

      This image was generated using ab220811, the same antibody but in a PBS-only buffer formulation. 

    • Anti-DUSP6 antibody [EPR129Y] (ab76310) at 1/5000 dilution (purified) + HepG2 cell lysate at 20 µg

      Secondary
      HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

      Predicted band size: 42 kDa
      Observed band size: 42,44 kDa
      why is the actual band size different from the predicted?



      Blocking buffer: 5% NFDM/TBST
      Dilution buffer: 5% NFDM/TBST

      ab76310 detects an unspecific band around 100 kDa in human materials.

    • Overlay histogram showing NIH-3T3 cells fixed in 4% PFA and stained with purified ab76310 at a dilution of 1 in 200 (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).
    • ab76310 (purified) at 1/20 immunoprecipitating DUSP6 in 10 μg NIH-3T3 (Lanes 1 and 2, observed at 42 and 44 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-DUSP6 antibody [EPR129Y] (ab76310) at 1/1000 dilution (purified)

      Lane 1 : rat brain lysate
      Lane 2 : NIH/3T3 cell lysate

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size: 42 kDa
      Observed band size: 42,44 kDa why is the actual band size different from the predicted?



      Blocking buffer: 5% NFDM/TBST
      Dilution buffer: 5% NFDM/TBST

    • Anti-DUSP6 antibody [EPR129Y] (ab76310) at 1/1000 dilution (purified) + mouse brain at 10 µg

      Secondary
      HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

      Predicted band size: 42 kDa
      Observed band size: 42,44 kDa why is the actual band size different from the predicted?



      Blocking buffer: 5% NFDM/TBST
      Dilution buffer: 5% NFDM/TBST

    • Anti-DUSP6 antibody [EPR129Y] (ab76310) at 1/500 dilution (unpurified) + 3T3 cell lysate at 10 µg

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

      Predicted band size: 42 kDa
      Observed band size: 42/44 kDa why is the actual band size different from the predicted?

    • All lanes : Anti-DUSP6 antibody [EPR129Y] (ab76310) at 1/4000 dilution (unpurified)

      Lane 1 : Marker
      Lane 2 : Lysate from wild type primary murine embryonic fibroblasts (MEFs) untreated at 10 µg
      Lane 3 : Lysate from wild type primary murine embryonic fibroblasts (MEFs) treated with 100ngµl-1, 12-O-tetradecanoylphorbol-13-acetate (TPA) for 2 hours at 10 µg
      Lane 4 : Lysate from MKP-3 null primary murine embryonic fibroblasts (MEFs) untreated at 10 µg
      Lane 5 : Lysate from MKP-3 null primary murine embryonic fibroblasts (MEFs) treated with 100ngµl-1, 12-O-tetradecanoylphorbol-13-acetate (TPA) for 2 hours at 10 µg

      Secondary
      All lanes : Goat anti Rabbit HRP conjugate at 1/10000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 42 kDa
      Observed band size: 42 kDa


      Exposure time: 2 minutes

      See Abreview

    • Immunohistochemistry (PFA perfusion fixed frozen sections) analysis of Mouse brain tissue sections labelling DUSP6 with unpurified ab76310 at 1/400 dilution for 14 hours at 4°C. A biotinylated polyclonal anti-rabbit IgG was used as the secondary antibody at 1/250 dilution.

      See Abreview

    • Immunohistochemical staining of paraffin-embedded human gastric carcinoma using unpurified ab76310 at 1/50 dilution.

    • Overlay histogram showing HeLa cells stained with unpurified ab76310 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76310, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde/permeabilized with 0.1% PBS-Tween 20 used under the same conditions.

    References

    This product has been referenced in:
    • Hu X  et al. Tripartite motif-containing protein 7 regulates hepatocellular carcinoma cell proliferation via the DUSP6/p38 pathway. Biochem Biophys Res Commun 511:889-895 (2019). Read more (PubMed: 30850165) »
    • Beaudry K  et al. Dual-specificity phosphatase 6 deletion protects the colonic epithelium against inflammation and promotes both proliferation and tumorigenesis. J Cell Physiol 234:6731-6745 (2019). Read more (PubMed: 30273442) »
    See all 19 Publications for this product

    Customer reviews and Q&As

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    1-6 of 6 Abreviews

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (skin)
    Permeabilization
    Yes - tween 0.05%
    Specification
    skin
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Acetone

    Abcam user community

    Verified customer

    Submitted Apr 29 2019

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Baboon Tissue sections (Prepubertal testis)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Sodium citrate
    Permeabilization
    No
    Specification
    Prepubertal testis
    Blocking step
    5% serum + 3% BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Feb 28 2017

    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Non-reduced Denaturing (10% SDS-PAGE)
    Sample
    Human Cell lysate - whole cell (Human lung cancer cells)
    Specification
    Human lung cancer cells
    Blocking step
    Odyssey Blocking Buffer in TBS-T as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 22°C

    Abcam user community

    Verified customer

    Submitted Oct 03 2014

    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Mouse Tissue sections (brain)
    Specification
    brain
    Fixative
    Formaldehyde
    Antigen retrieval step
    None
    Permeabilization
    Yes - 0.1% Tween-20
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Temperature: 24°C

    Dr. Carlos Sindreu

    Verified customer

    Submitted May 10 2013

    This product is known to not work in this application or species.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Cultured Myofiber)
    Specification
    Cultured Myofiber
    Fixative
    Formaldehyde
    Permeabilization
    Yes - 0,2% Triton
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

    Fabien Le Grand

    Verified customer

    Submitted Nov 07 2012

    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Primary MEF)
    Loading amount
    10 µg
    Specification
    Primary MEF
    Treatment
    100 ng/µl TPA for 2 hours
    Gel Running Conditions
    Reduced Denaturing (4-14% Bis-Tris)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

    Graeme Stewart

    Verified customer

    Submitted Jul 24 2012

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