Recombinant
RabMAb

Anti-DUSP6 antibody [EPR129Y] (ab76310)

Overview

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

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human DUSP6 aa 350 to the C-terminus (C terminal).
    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.

ICC/IF 1/200.

For unpurified, use 1/1000.

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

  • 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.

  • 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
    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
    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

    This image was taken from an abreview by Graeme Stewart.

    See Abreview

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

  • 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

  • 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.

  • ICC/IF image of unpurified ab76310 stained PC12 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 (ab76310, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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.

  • 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
  • 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).
  • 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:
  • Zhou J  et al. ROS-mediated Different Homeostasis of Murine Corneal Epithelial Progenitor Cell Line under Oxidative Stress. Sci Rep 6:36481 (2016). WB . Read more (PubMed: 27805062) »
  • Schaffert SA  et al. mir-181a-1/b-1 Modulates Tolerance through Opposing Activities in Selection and Peripheral T Cell Function. J Immunol 195:1470-9 (2015). Read more (PubMed: 26163591) »

See all 11 Publications for this product

Customer reviews and Q&As

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
Username

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
Username

Abcam user community

Verified customer

Submitted Oct 03 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Dr. Carlos Sindreu

Verified customer

Submitted May 10 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Fabien Le Grand

Verified customer

Submitted Nov 07 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Graeme Stewart

Verified customer

Submitted Jul 24 2012

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

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