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Anti-DUSP6 Antibody EPR129Y
Also for DUSP6 (NM_022652)
|A synthetic peptide corresponding to residues near the C-terminus of human MKP-3 was used as an immunogen.|
|Human, Mouse, Rat
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, IF, FC
||IHC-P: 1:50 - 1:100; IHC-FoFr: Use at an assay dependent concentration; WB: 1:500; IP: 1:40; FC: 1:20 - 1:60; ICC/IF: 1:1000
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant (Protein A or G Sepharose)
|Is unsuitable for ICC.
|Homo sapiens dual specificity phosphatase 6 (DUSP6), transcript variant 2|
|HH19; MKP3; PYST1|
|MKP-3 is expressed constitutively in human skin fibroblasts and dephosphorylates and inactivates MAP kinase in vitro and in vivo. MKP-3 displays very low activity towards the stress-activated protein kinases (SAPKs) or RK/p38 in vitro, indicating that these kinases are not physiological substrates for MKP-3. MKP-3 is thought to play a role in the dephosphorylation and inactivation of MAP kinases and are therefore likely to be important in the regulation of diverse cellular processes such as proliferation, differentiation, and apoptosis. For this reason it has been suggested that MAP kinase phosphatases may be tumor suppressors (1). MKP-3 is expressed in lung, heart, brain, and kidney, but not significantly in skeletal muscle or testis. In situ hybridization studies of MKP-3 in brain reveal enrichment within the CA1, CA3, and CA4 layers of the hippocampus. Observations indicate that MKP-3 is a novel dual-specificity phosphatase that displays a distinct tissue distribution, subcellular localization, and regulated expression, suggesting a unique function in controlling MAP kinase family members (2). |
MAPK signaling pathway
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Western blot - Anti-DUSP6 antibody [EPR129Y]; All lanes : Anti-DUSP6 antibody [EPR129Y] at 1/4000 dilution.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 minutesThis image was taken from an abreview by Graeme Stewart.
Western blot - DUSP6 antibody [EPR129Y]; Anti-DUSP6 antibody [EPR129Y] at 1/500 dilution + 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 .
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-DUSP6 antibody [EPR129Y]; Immunohistochemistry (PFA perfusion fixed frozen sections) analysis of Mouse brain tissue sections labelling DUSP6 with 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - DUSP6 antibody [EPR129Y]; Immunohistochemical staining of paraffin-embedded human gastric carcinoma using ab76310 at 1/50 dilution.
Immunocytochemistry/ Immunofluorescence - DUSP6 antibody [EPR129Y]; ICC/IF image of 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 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.
Flow Cytometry-DUSP6 antibody [EPR129Y](ab76310); Overlay histogram showing HeLa cells stained with 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 for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x10^6 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.