Anti-MEK3 (phospho S189/T193) + MEK6 (phospho S207/T211)抗体
兔多克隆抗体to MEK3 (phospho S189/T193) + MEK6 (phospho S207/T211)
Rabbit
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.
适用于: WBmore details
与反应: Human
预测可用于: a wide range of other species
Synthetic peptide corresponding to Human MEK3 (phospho S189/T193) + MEK6 (phospho S207/T211).
Mal-E-tagged fusion protein expressing MEK 6, left inactivated or activated by adding MEKK, Anisomycin treated A549 cell line
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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Liquid
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 0.1% BSA
BSA is IgG and protease free
Immunogen affinity purified
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.
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.
多克隆
IgG
Abpromise™承诺保证使用ab4759于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB | 1/500 - 1/2000. Predicted molecular weight: 39.3 kDa. |
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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 kDawhy 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
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.
