Anti-MEK3 (phospho S189/T193) + MEK6 (phospho S207/T211) antibody (ab4759)
Key features and details
- Rabbit polyclonal to MEK3 (phospho S189/T193) + MEK6 (phospho S207/T211)
- Suitable for: WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-MEK3 (phospho S189/T193) + MEK6 (phospho S207/T211) antibody -
Description
Rabbit polyclonal to MEK3 (phospho S189/T193) + MEK6 (phospho S207/T211) -
Host species
Rabbit -
Specificity
This antibody reacts with MEK 3 and MEK 6 (93% homologous). This antibody shows some cross-reactivity with MEK 4 when tested in a system with high MEK 4 expression levels. It does not react with any other MEK isoforms. -
Tested applications
Suitable for: WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: a wide range of other species
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Immunogen
Synthetic peptide corresponding to Human MEK3 (phospho S189/T193) + MEK6 (phospho S207/T211).
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Positive control
- Mal-E-tagged fusion protein expressing MEK 6, left inactivated or activated by adding MEKK, Anisomycin treated A549 cell line
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General notes
Mitogen Activated Protein Kinase Kinases 3 and 6 (MEK 3/6 or MKK 3/6) are 42 kDa members of a tyrosine/threonine protein kinase family that activate p38, which is part of the inflammation/stress signaling pathway. Phosphorylation of MEK3 and 6 by MEKK1 on serine 189 and threonine 193 (serine 207 and threonine 211 for MEK6) in the catalytic domain activates the proteins and enables them to phosphorylate p38.
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Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 0.1% BSA
BSA is IgG and protease free -
Concentration information loading... -
Purity
Immunogen affinity purified -
Purification notes
Purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated MEK 3. The final product is generated by affinity chromatography using a MEK 3 derived peptide that is phosphorylated at serine 189 and threonine 193. -
Primary antibody notes
Mitogen Activated Protein Kinase Kinases 3 and 6 (MEK 3/6 or MKK 3/6) are 42 kDa members of a tyrosine/threonine protein kinase family that activate p38, which is part of the inflammation/stress signaling pathway. Phosphorylation of MEK3 and 6 by MEKK1 on serine 189 and threonine 193 (serine 207 and threonine 211 for MEK6) in the catalytic domain activates the proteins and enables them to phosphorylate p38. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab4759 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 |
1/500 - 1/2000. Predicted molecular weight: 39.3 kDa.
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| Notes |
|---|
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WB
1/500 - 1/2000. Predicted molecular weight: 39.3 kDa. |
Target
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Alternative names
- AW212142 antibody
- dual specificity mitogen activated protein kinase kinase 3 antibody
- Dual specificity mitogen activated protein kinase kinase 6 antibody
see all
Images
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Western blot - Anti-MEK3 (phospho S189/T193) + MEK6 (phospho S207/T211) antibody (ab4759)All lanes : Anti-MEK3 (phospho S189/T193) + MEK6 (phospho S207/T211) antibody (ab4759) at 1/2000 dilution
Lane 1 : A549 whole cell extract
Lane 2 : A549 treated for 20 minutes with 200 nM of PMA
Lane 3 : A549 treated for 30 minutes with 25 µg/mL of Anisomycin
Lane 4 : A549 treated for 40 minutes with UV
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/2500 dilution
Predicted band size: 39.3 kDa
Observed band size: 39, 37 kDa why is the actual band size different from the predicted?Two bands ~ 37 and 39 kDa corresponding to MEK3 (pS189/pT193) + MEK6 (pS207/pT211) was observed in Anisomycin treated cell line tested
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Western blot - Anti-MEK3 (phospho S189/T193) + MEK6 (phospho S207/T211) antibody (ab4759)Peptide Competition: Extracts prepared from background extracts with Mal-E tagged fusion protein expressing MEK 6 left inactivated (6) or with MEKK added for activation (1-5) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4oC, then were incubated with 0.50 µg/mL ab4759 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 6), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic hosphoserine-containing peptide (3), a generic phosphothreonine-containing peptide (4), or the phosphopeptide immunogen (5). After washing, the membrane was incubated with goat F(ab"e;)2 anti-rabbit IgG alkaline phosphatase and signals were detected. The data show that only the phosphopeptide corresponding to MEK3/6 [pSpT189/193]/[pSpT207/211] blocks the antibody signal, demonstrating the specificity of the antibody. Please note that the Mal-E-tagged fusion protein expressing MEK6 runs at ~81 kDa.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab4759 has not yet been referenced specifically in any publications.