Anti-MEK1 (phospho T292) antibody (ab5612)
Key features and details
- Rabbit polyclonal to MEK1 (phospho T292)
- Suitable for: WB
- Reacts with: Recombinant fragment
- Isotype: IgG
Overview
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Product name
Anti-MEK1 (phospho T292) antibody
See all MEK1 primary antibodies -
Description
Rabbit polyclonal to MEK1 (phospho T292) -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species WB Recombinant fragment
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Immunogen
The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human MEK 1 that contains threonine 292.
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Positive control
- NIH3T3 cells +/- PDGF.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol (glycerin, glycerine), 0.1% BSA -
Concentration information loading... -
Purity
Immunogen affinity purified -
Purification notes
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. The final product is generated by affinity chromatography using a MEK 1 derived peptide that is phosphorylated at threonine 292. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-MEK1 (phospho T292) antibody (ab5612)Peptide Competition and Mutant Analysis: Recombinant wild-type (lanes 1 4) and mutant (T292A) MEK1 (lane 5) treated with ERK to induce phosphorylation was added to background extracts and resolved by SDS PAGE on a 10% Tris-glycine gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4oC, then were incubated with 0.50
µ g/mL ab5612 antibody for two hours at room temperature, following prior incubation with: no peptide (1, 5), the nonphosphopeptide corresponding to the immunogen (2), a generic phosphothreonine containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F(ab’ 2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to ab5612 blocks the antibody signal, and the T292A mutant is not reactive, demonstrating the specificity of the antibody. The recombinant wild-type and mutant MEK1 were kindly provided by Dr. Natalie Ahn, University of Colorado.
Peptide Competition and Mutant Analysis: Recombinant wild-type (lanes 1 4) and mutant (T292A) MEK1 (lane 5) treated with ERK to induce phosphorylation was added to background extracts and resolved by SDS PAGE on a 10% Tris-glycine 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 ab5612 antibody for two hours at room temperature, following prior incubation with: no peptide (1, 5), the nonphosphopeptide corresponding to the immunogen (2), a generic phosphothreonine containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F(ab’ 2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to ab5612 blocks the antibody signal, and the T292A mutant is not reactive, demonstrating the specificity of the antibody. The recombinant wild-type and mutant MEK1 were kindly provided by Dr. Natalie Ahn, University of Colorado.
