Recombinant
RabMAb

Recombinant Anti-DUSP6 antibody [EPR129Y] - BSA and Azide free (ab220811)

Overview

  • Product name

    Anti-DUSP6 antibody [EPR129Y] - BSA and Azide free
    See all DUSP6 primary antibodies
  • Description

    Rabbit monoclonal [EPR129Y] to DUSP6 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IF, IHC-FoFrmore 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

    Ab220811 is the carrier-free version of ab76310. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab220811 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

    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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR129Y
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab220811 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 Use at an assay dependent concentration. Detects a band of approximately 42,44 kDa (predicted molecular weight: 42 kDa).Can be blocked with DUSP6 peptide (ab171765).
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration.

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

  • 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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76310).

  • 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76310).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76310).

  • 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.

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76310).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76310).

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76310).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76310).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76310).

References

This product has been referenced in:

  • Schmidt C  et al. Immunoproteasome Inhibition Impairs T and B Cell Activation by Restraining ERK Signaling and Proteostasis. Front Immunol 9:2386 (2018). IP, WB ; Mouse . Read more (PubMed: 30416500) »
  • 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 9 Publications for this product

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