Application
A549 cells plated on fibronectin, serum starved overnight then stimulated with serum for 1 hour. Green = MEK1 [pS298] Red = Actin Blue = Hoechst 33342Application
Extracts of NIH3T3 cells untreated (1) or treated with 50 ng/mL PDGF for 15 minutes (2-7) were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was either left untreated (1-5, 7) or treated with lambda phosphatase (6), blocked with a 4% BSA-TBST buffer for one hour at room temperature, then incubated with the MEK1 [pS298] antibody for two hours at room temperature in a 1% BSA-TBST, following prior incubation with: no peptide (1, 2, 6, 7), the non-phosphopeptide corresponding to the phosphopeptide immunogen (3), a generic phosphoserine-containing peptide (4) or the phosphopeptide immunogen (5). After washing, the membrane was incubated with goat F(ab)2 anti-rabbit IgG HRP conjugate and signals were detected. The data show that only the phosphopeptide corresponding to MEK1 [pS298] blocks the antibody signal, demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.| Product Name | MEK1 (phospho Ser298) antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Rabbit polyclonal to MEK1 ( phospho S298 )Mitogen activated protein kinase kinase 2 (MEK 2 or MAPKK 2) is a member of a family of tyrosine/threonine protein kinases that activate the ERK 1 and 2/ MAPK enzymes by phosphorylating both residues within the threonine - glutamate - tyrosine (TEY) motif in the activation loop. MEK 1 and 2 are also activated by dual phosphorylation, which occurs on serine 218 and 222, in the activation loop of the MEK. Serine 298 of MEK1 is phosphorylated by PAK 1, which promotes MEK 1 binding to c Raf and its subsequent phosphorylation of MEK 1. |
| Immunogen | The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human MEK 1 that contains serine 298. |
| Modification | p-Ser298 |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFWB |
| WB: Use at 1/1000. Detects a band of approximately 50 kDa. Optimal dilutions/concentrations should be determined by the end user.: |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 0.5mg/ml |
| Purification | Affinity purified |
| Gene Synonyms | MAPKK1 MEK1 MKK1 PRKMK1 MEK1 (phospho S298) MEK1 (phospho Ser298) MEK1 (pS298) MEK1 (pSer298) MEK1 phospho S298 MEK1 phospho Ser298 Phospho MEK1 (S298) Phospho MEK1 (Ser298) Phospho MEK1 (pS298) Phospho MEK1 (pSer298) |
|---|---|
| Target | MEK1 |
| Cellular Localization | Cytoplasmic |
Application
A549 cells plated on fibronectin, serum starved overnight then stimulated with serum for 1 hour. Green = MEK1 [pS298] Red = Actin Blue = Hoechst 33342
Application
Extracts of NIH3T3 cells untreated (1) or treated with 50 ng/mL PDGF for 15 minutes (2-7) were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was either left untreated (1-5, 7) or treated with lambda phosphatase (6), blocked with a 4% BSA-TBST buffer for one hour at room temperature, then incubated with the MEK1 [pS298] antibody for two hours at room temperature in a 1% BSA-TBST, following prior incubation with: no peptide (1, 2, 6, 7), the non-phosphopeptide corresponding to the phosphopeptide immunogen (3), a generic phosphoserine-containing peptide (4) or the phosphopeptide immunogen (5). After washing, the membrane was incubated with goat F(ab)2 anti-rabbit IgG HRP conjugate and signals were detected. The data show that only the phosphopeptide corresponding to MEK1 [pS298] blocks the antibody signal, demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.| Positive Control | NIH3T3 cells +/- PDGF, A431 +/- EGF, PC12 +/- NGF |
|---|---|
| Application Notes | WB: Use at 1/1000. Detects a band of approximately 50 kDa. Optimal dilutions/concentrations should be determined by the end user.: |
| Form | Liquid |
|---|---|
| Storage Instructions | Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | Preservative: 0.05% Sodium Azide; Constituents: PBS, 50% glycerol, 1mg/ml BSA (IgG, protease free). pH 7.3 |
