Anti-MEK1 + MEK2 (phospho S222)抗体
参阅全部 MEK1 + MEK2 一抗
兔多克隆抗体to MEK1 + MEK2 (phospho S222)
Rabbit
适用于: ICC/IF, WBmore details
与反应: Mouse, Human
预测可用于: Chicken, Xenopus laevis, Chimpanzee
Synthetic peptide corresponding to Human MEK1 + MEK2 (phospho S222).
ICC/IF: Mouse embryonic fibroblasts WB: NIH3T3 cells
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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.
Preservative: 0.05% Sodium azide
Constituents: PBS, 0.1% BSA
BSA is IgG and protease free
浓度
50 µl 浓度为 0.5 mg/ml
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 1 + 2. The final product is generated by affinity chromatography using a MEK 1 + 2 derived peptide that is phosphorylated at serine 222.
多克隆
IgG
Abpromise™承诺保证使用ab4750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 |
|---|---|---|
| ICC/IF | Use at an assay dependent concentration. Assay dependent. | |
| WB | 1/1000. Predicted molecular weight: 43.3 kDa. |
Entrez Gene: 5604 Human
Entrez Gene: 5605 Human
Entrez Gene: 26395 Mouse
Entrez Gene: 26396 Mouse
Entrez Gene: 379991 Xenopus laevis
Omim: 176872 Human
Omim: 601263 Human
SwissProt: Q90891 Chicken
SwissProt: P36507 Human
SwissProt: Q02750 Human
SwissProt: P31938 Mouse
SwissProt: Q63932 Mouse
SwissProt: Q05116 Xenopus laevis
Unigene: 145442 Human
Unigene: 248907 Mouse
Unigene: 5877 Xenopus laevis
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Western blot - Anti-MEK1 + MEK2 (phospho S222) antibody (ab4750)
All lanes : Anti-MEK1 + MEK2 (phospho S222) antibody (ab4750)
Lane 1 : A549 whole cell extract lysate
Lane 2 : A549 whole cell lysate treated with EGF (200 ng/mL for 10 min)
Lane 3 : A549 whole cell lysate treated with Afatinib (0.5 uM for 16 h) followed by EGF (200 ng/mL for 10 min)
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate at 0.25 µg/ml
Predicted band size: 43.3 kDa
Western blot analysis was performed on whole cell extracts of A549 in different conditions, observed a 45 kDa band corresponding to Phospho-MEK1/MEK2 (Ser222) was observed in untreated A549 lysate, the signal increased upon EGF treatment, and decreased upon pretreatment with the EGFR antagonist, Afatinib. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel, XCell SureLock™ Electrophoresis System and Novex® Sharp Pre-Stained Protein Standard. Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate.
Western blot - Anti-MEK1 + MEK2 (phospho S222) antibody (ab4750)
Lane 1 : Anti-MEK1 + MEK2 (phospho S222) antibody (ab4750)
Lane 1 : Untreated NIH3T3 cells
Lanes 2-5 : Anti-MEK1 + MEK2 (phospho S222) antibody (ab4750)
Secondary
Lane 1 : A goat F (ab)2 anti-rabbit IgG alkaline phosphatase
Predicted band size: 43.3 kDa
Peptide Competition and Stimulation Extracts of NIH3T3 cells untreated (lane 1) or treated with 50 ng/mL PDGF for 15 minutes (2-5) were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 4% BSA-TBST buffer overnight at 4°C, then incubated with the MEK1&2 (pS222) antibody for two hours at room temperature in a 1% BSA-TBST buffer, following various prior incubation conditions. After washing, the membrane was incubated with goat F (ab)2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Pierce SuperSignal method. The data show that only the phosphopeptide corresponding to MEK1&2 (pS222) block the antibody signal, demonstrating the specificity of the antibody. The data also show the induction of MEK1&2 (pS222) phosphorylation by the addition of PDGF to this cell system.
Immunocytochemistry/ Immunofluorescence - Anti-MEK1 + MEK2 (phospho S222) antibody (ab4750)
Immunofluorescent analysis of mouse embryonic fibroblasts labelled for MEK1/2 (pS222) using ab4750, phosphospecific antibody. The cell is actively dividing. Blue represents chromosomes in anaphase of mitotic cell division. Green shows mitotic spindle expressing phosphorylated MEK1/2.
