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HTRF Human Phospho-MEK1 (Ser298) Detection Kit, 10,000 Assay points

This HTRF kit enables the cell-based quantitative detection of MEK1 phosphorylation at Ser298.

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption & disposal requirements under European REACH regulations (EC 1907/2006).
Part Number: 64MEK1S98PEY
Unit Size: 50,000 Assay Points
Part Number: 64MEK1S98PEG
Unit Size: 500 Assay Points
Part Number: 64MEK1S98PEH
Unit Size: 10,000 Assay Points
  • The Phospho-MEK1 (Ser298) kit measures endogenous MEK1 protein phophorylated at Ser298, to help studies of 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.

  • Application
    Protein Detection
    Assay Points
    10000
    Assay Target Type
    Kit
    Assay Technology
    HTRF
    Automation Compatible
    Yes
    Brand
    HTRF
    Detection Method
    HTRF
    Experimental Type
    In vitro
    Therapeutic Area
    Cardiovascular
    Neuroscience
    Oncology & Inflammation
    Unit Size
    10,000 Assay Points
  • Phospho-MEK1 (Ser298) assay principle

    The Phospho-MEK1 (Ser298) assay measures MEK1 when phosphorylated at Ser298. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Phospho-MEK1 (Ser298) assay uses two antibodies: one labeled with a donor fluorophore, the other one with an acceptor. The first antibody was 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 protein’s phosphorylation state under a no-wash assay format.

    Phospho how it works phospho mek1 ser298 cellular assay principle
    Phospho-MEK1 (Ser298) 2-plate assay protocol

    The 2 plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a 384-well low volume detection plate before the addition of Phospho-MEK1 (Ser298) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

    Phospho how it works phospho mek1 ser298 2 plate assay protocol
    Phospho-MEK1 (Ser298) 1-plate assay protocol

    Detection of Phosphorylated MEK1 (Ser298) 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.

    Phospho how it works total mek 1 plate assay protocol

     

  • Detection of phospho MEK1 (Ser298) on Hela cells

    Hela cells were cultured in a T175 flask. At 80% of confluency, the cell culture medium was removed and 3mL of complete lysis buffer (1X) were added. The lysis step was done under gentle shaking for 30 minutes. Lysate was then collected and diluted. 16µL of each dilution were transferred into a 384-well plate for the detection of phospho MEK1 (Ser298).

    Detection of phospho MEK1 on Hela cells

     

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Guide
HTRF solutions, guide to major applications

This guide provides you an overview of HTRF applications in several therapeutic areas.