Phospho-MEK1 (Ser298) cellular kit
All-in-one kit for robust detection of Phospho-MEK1 (Ser298)
Phospho-MEK1 (Ser218/222) cellular kit HTRF®
The Phospho-MEK1 (Ser218/222) kit enables the cell-based detection of MEK1 protein phosphorylated at Ser218/222 when studying the RAS-RAF-MEK cascade in the MAPK/ERK pathway.
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High sensitivity
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Faster and more convenient than ELISA
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Ready-to-use
Overview
The Phospho-MEK1 (Ser218/222) kit measures endogenous MEK1 protein phosphorylated at Ser218/222 when studying the RAS-RAF-MEK cascade in the MAPK/ERK pathway. Phosphorylated by RAF, this dual specificity mitogen-activated protein kinase is responsible for the phosphorylation of ERK, one of the most important signaling nodes, and protein kinase in cell proliferation, survival, and differentiation pathways.
Benefits
- SPECIFICITY
- PRECISION
Phospho-MEK1 (Ser218/222) assay principle
The Phospho-MEK1 (Ser218/222) assay measures MEK1 when phosphorylated at Ser218/222. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Phospho-MEK1 (Ser218/222) assay uses 2 labeled antibodies: one with a donor fluorophore, the other one with an acceptor. The first antibody is selected for its specific binding to the phosphorylated motif on the protein, the second for its ability to recognize the protein independent of its phosphorylation state. Protein phosphorylation enables an immune-complex formation involving both labeled antibodies and which brings the donor fluorophore into close proximity to the acceptor, thereby generating a FRET signal. Its intensity is directly proportional to the concentration of phosphorylated protein present in the sample, and provides a means of assessing the proteins phosphorylation state under a no-wash assay format.
Phospho-MEK1 (Ser218/222) 2-plate assay protocol
The 2 plate protocol involves culturing cells in a 96-well plate before lysis then transferring lysates to a 384-well low volume detection plate before adding Phospho-MEK1 (Ser218/222) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.
Phospho-MEK1 (Ser218/222) 1-plate assay protocol
Detection of Phosphorylated MEK1 (Ser218/222) with HTRF reagents can be performed in a single plate used for culturing, stimulation and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality.
WB versus HTRF assay for phospho-MEK1 (Ser218/222)
HeLa cells were grown in a T175 flask 37°C, 5% Co2, 2 days. Stimulation was done with EGF 2nM for 5 min. After removal of cell culture medium, 3 mL of supplemented lysis buffer were added and incubated for 30 min. Soluble supernatants were collected after 10 min centrifuging. Equal amounts of lysates were used for a side by side comparison of WB and HTRF.
Phospho-MEK1 assay shows the same level of sensitivity as Western Blot.
PMA dose-response on NIH 3T3 cells
Murine NIH 3T3 cells (25,000 cells/well) were stimulated for 15 minutes at 37°C with various concentrations of PMA.
After a 30 minutess lysis incubation time, phosphorylated MEK1 were measured using the two-plate assay protocol.
Inhibition effect of EGRF & RAF inhibitors on HeLa cells
HeLa cells (50,000 cells/well) were incubated for 15 min at 37°C with various concentrations of the 2 inhibitors, Erlotinib (the EGFR inhibitor) and L777,450 (a Raf inhibitor). EGF 0.8nM (agonist) was then added and incubated for 10 minutes.
After a 30 minutess lysis incubation time, inhibition of the phosphorylated MEK was measured using the Phospho-MEK1 assay. Assay was done using the two-plate assay protocol.
Simplified pathway dissection with HTRF phospho-assays and CyBi-felix liquid handling
Analyse of PI3K/AKT/mTor translational control pathway - Application Notes
Investigating kinase activity in a cellular context
HTRF cellular assays - Scientific Presentations
Open R&D: Sanofi Access Platform
In collaboration with Sanofi - Scientific Presentations
Cisbio lysis buffer compatibility
Cell Signaling: Biomarkers, Phospho- & total-protein Assays - Flyers
HTRF cellular phospho-protein assays
Physiologically relevant results fo fast flowing research - Flyers
HTRF products and supporting scientific content to investifgate GPCRs - Flyers
Cell Signaling: Biomarkers, Phospho- & total-protein assays - Flyers
HTRF assays for Oncology and Inflammation
Signaling in the immune system - Brochures
PI3K/AKT/mTor translational control pathway - Posters
Universal HTRF® phospho-protein platform: from 2D, 3D, primary cells to patient derived tumor cells
Analysis of a large panel of diverse biological samples and cellular models - Posters
Multidimentional GPCR investigations - Posters
From 2D, 3D cell cultures to xenografts: A smart HTRF platform to maximize anticancer drug discovery
One technology across all samples - Application Notes
HTRF phospho assays reveal subtle drug induced effects in tumor-xenografts
Tumor xenograft analysis: HTRF versus Western blot - Application Notes
HTRF cell-based phospho-protein data normalization
Valuable guidelines for efficiently analyzing and interpreting results - Application Notes
HTRF phospho-total lysis buffer: a universal alternative to RIPA lysis buffers
Increased flexibility of phospho-assays - Application Notes
HTRF and Tag-lite technologies in multidimensional cell-based assays for GPCR investigations
16 GPCR challenges, 1 solution - Posters
Multi-tissue cellular modeling and anlysis of insulin signaling - Posters
Best practices for analyzing brain samples with HTRF® phospho assays for neurosciences
Insider Tips for successful sample treatment - Technical Notes
HTRF Alpha-tubulin Housekeeping kit
Properly interpret your compound effect - Application Notes
Understanding GPCRs is the key to improved DD
In collaboration with GEN - Guides
Optimize your HTRF cell signaling assays on tissues
HTRF and WB compatible guidelines - Technical Notes
Key guidelines to successful cell signaling experiments
Mastering the art of cell signaling assays optimization - Guides
HTRF phospho-assays reveal subtle drug-induced effects
Detailed protocol and direct comparison with WB - Posters
A single technology for 2D cells, 3D cells, and xenograft models - Posters
Best practices for analyzing tumor xenografts with HTRF phospho assays
Protocol for tumor xenograft analysis with HTRF - Technical Notes
How to run a cell based phospho HTRF assay
What to expect at the bench - Videos
Unleash the potential of your phosphorylation research with HTRF
Unmatched ease of use, sensitivity and specificity assays - Videos
Cell-based kinase assays in HTS ? potential and limitations for primary and secondary screening
In collaboration with Bayer - Scientific Presentations
Product Insert MEK1 P-S218/222 Kit / 64ME1PEG-64ME1PEH
64ME1PEG-64ME1PEH - Product Insert
All your HTRF assays in one document! - Catalog
A guide to Homogeneous Time Resolved Fluorescence
General principles of HTRF - Guides
How HTRF compares to Western Blot and ELISA
Get the brochure about technology comparison. - Brochures
HTRF® cell signaling platform combined with iCell® Hepatocytes
A solution for phospho-protein analysis in metabolic disorders - Posters
Unleash the potential of your phosphorylation research with HTRF
A fun video introducing you to phosphorylation assays with HTRF - Videos
How to run a cell based phospho HTRF assay
3' video to set up your Phospho assay - Videos
Safety Data Sheet (DEU) MEK1 P-S218/222 Kit / 64ME1PEG
64ME1PEG - Safety Data Sheet
Safety Data Sheet (ELL) MEK1 P-S218/222 Kit / 64ME1PEG
64ME1PEG - Safety Data Sheet
Safety Data Sheet (FRA-FR) MEK1 P-S218/222 Kit / 64ME1PEG
64ME1PEG - Safety Data Sheet
Safety Data Sheet (ITA) MEK1 P-S218/222 Kit / 64ME1PEG
64ME1PEG - Safety Data Sheet
Safety Data Sheet (SPA) MEK1 P-S218/222 Kit / 64ME1PEG
64ME1PEG - Safety Data Sheet
Safety Data Sheet (ENG-GB) MEK1 P-S218/222 Kit / 64ME1PEG
64ME1PEG - Safety Data Sheet
Safety Data Sheet (ENG-US) MEK1 P-S218/222 Kit / 64ME1PEG
64ME1PEG - Safety Data Sheet
Safety Data Sheet (DEU) MEK1 P-S218/222 Kit / 64ME1PEH
64ME1PEH - Safety Data Sheet
Safety Data Sheet (ELL) MEK1 P-S218/222 Kit / 64ME1PEH
64ME1PEH - Safety Data Sheet
Safety Data Sheet (FRA-FR) MEK1 P-S218/222 Kit / 64ME1PEH
64ME1PEH - Safety Data Sheet
Safety Data Sheet (ITA) MEK1 P-S218/222 Kit / 64ME1PEH
64ME1PEH - Safety Data Sheet
Safety Data Sheet (SPA) MEK1 P-S218/222 Kit / 64ME1PEH
64ME1PEH - Safety Data Sheet
Safety Data Sheet (ENG-GB) MEK1 P-S218/222 Kit / 64ME1PEH
64ME1PEH - Safety Data Sheet
Safety Data Sheet (ENG-US) MEK1 P-S218/222 Kit / 64ME1PEH
64ME1PEH - Safety Data Sheet
Safety Data Sheet (DEU) MEK1 P-S218/222 Kit / 64ME1PEY
64ME1PEY - Safety Data Sheet
Safety Data Sheet (ELL) MEK1 P-S218/222 Kit / 64ME1PEY
64ME1PEY - Safety Data Sheet
Safety Data Sheet (FRA-FR) MEK1 P-S218/222 Kit / 64ME1PEY
64ME1PEY - Safety Data Sheet
Safety Data Sheet (ITA) MEK1 P-S218/222 Kit / 64ME1PEY
64ME1PEY - Safety Data Sheet
Safety Data Sheet (SPA) MEK1 P-S218/222 Kit / 64ME1PEY
64ME1PEY - Safety Data Sheet
Safety Data Sheet (ENG-GB) MEK1 P-S218/222 Kit / 64ME1PEY
64ME1PEY - Safety Data Sheet
Safety Data Sheet (ENG-US) MEK1 P-S218/222 Kit / 64ME1PEY
64ME1PEY - Safety Data Sheet
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