Recombinant Anti-DUSP6 antibody [EPR129Y] (ab76310)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR129Y] to DUSP6
- Suitable for: IHC-P, Flow Cyt (Intra), WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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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, Flow Cyt (Intra), WB, IPmore details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
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Positive control
- WB: 3T3, and A431 cell lysates; IHC-P: Human gastric carcinoma and human pancreas tissues; IHC-Fr: Mouse brain tissue; Flow Cyt (intra): NIH-3T3 and HeLa cells; IP: NIH-3T3 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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.
Properties
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR129Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Immunizing Peptide (Blocking)
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
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IHC-P | (1) |
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/20 - 1/200.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (8) |
1/500 - 1/1000. Detects a band of approximately 42,44 kDa (predicted molecular weight: 42 kDa).Can be blocked with DUSP6 peptide (ab171765).
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IP | (2) |
1/40 - 1/50.
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Notes |
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IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/20 - 1/200. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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. |
Target
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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
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Database links
- Entrez Gene: 1848 Human
- Entrez Gene: 67603 Mouse
- Entrez Gene: 116663 Rat
- Omim: 602748 Human
- SwissProt: Q16828 Human
- SwissProt: Q9DBB1 Mouse
- SwissProt: Q64346 Rat
- Unigene: 298654 Human
see all -
Alternative names
- Dual specificity phosphatase 6 antibody
- Dual specificity phosphatase 6 isoform a antibody
- Dual specificity protein phosphatase 6 antibody
see all
Images
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All lanes : Anti-DUSP6 antibody [EPR129Y] (ab76310) at 1/500 dilution
Lane 1 : Wild-type A431 whole cell lysate
Lane 2 : DUSP6 knockout A431 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 42 kDaLanes 1 - 3: Merged signal (red and green). Green - ab76310 observed at 42 kDa. Red - loading control, ab130007, observed at 130 kDa.
ab76310 was shown to recognize in wild-type A431 cells as signal was lost at the expected MW in DUSP6 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and DUSP6 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab76310 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DUSP6 antibody [EPR129Y] (ab76310)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.
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Western blot - Anti-DUSP6 antibody [EPR129Y] (ab76310)Schmidt C et al., Front Immunol, 9, 2386, 2018 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
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.
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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/TBSTab76310 detects an unspecific band around 100 kDa in human materials.
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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).
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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
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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? -
Western blot - Anti-DUSP6 antibody [EPR129Y] (ab76310)This image was taken from an abreview by Graeme Stewart.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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DUSP6 antibody [EPR129Y] (ab76310)
Immunohistochemical staining of paraffin-embedded human gastric carcinoma using unpurified ab76310 at 1/50 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Overlay histogram showing HeLa cells stained with unpurifiedab76310 (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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (37)
ab76310 has been referenced in 37 publications.
- Ye Z et al. The GSK3ß-ß-catenin-TCF1 pathway improves naive T cell activation in old adults by upregulating miR-181a. NPJ Aging Mech Dis 7:4 (2021). PubMed: 33558531
- Zewdu R et al. An NKX2-1/ERK/WNT feedback loop modulates gastric identity and response to targeted therapy in lung adenocarcinoma. Elife 10:N/A (2021). PubMed: 33821796
- Fedele C et al. SHP2 inhibition diminishes KRASG12C cycling and promotes tumor microenvironment remodeling. J Exp Med 218:N/A (2021). PubMed: 33045063
- Li Y et al. Clostridium difficile toxin B induces colonic inflammation through the TRIM46/DUSP1/MAPKs and NF-?B signalling pathway. Artif Cells Nanomed Biotechnol 48:452-462 (2020). PubMed: 31918570
- Zhu H et al. Knockdown of TRIM37 Promotes Apoptosis and Suppresses Tumor Growth in Gastric Cancer by Inactivation of the ERK1/2 Pathway. Onco Targets Ther 13:5479-5491 (2020). PubMed: 32606764