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    Phospho-MEK1 (S217/S221) ELISA Kit

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    2 Citations

    RayBio® Human/Mouse/Rat Phospho-MEK1 (S217/S221) ELISA Kit. This assay semi-quantitatively measures phosphorylated Mek (Ser217/S221) in lysate samples.

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    Catalog #:
    PEL-Mek-S217
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    Product Description

    Specifications

    Size 1 Plate Kit, 2 Plate Kit, 5 Plate Kit
    Species Human, Mouse, Rat
    Accession Number Q02750
    Gene Id 5604
    Gene Symbols MAP2K1|MEK1|PRKMK1
    Protein Name / Synonyms Dual specificity mitogen-activated protein kinase kinase 1 (MAP kinase kinase 1) (MAPKK 1) (MKK1) (EC 2.7.12.2) (ERK activator kinase 1) (MAPK/ERK kinase 1) (MEK 1)
    Quantitative/Semi-Quantitative Semi-Quantitative
    Specificity The antibody pair provided in this kit recognizes human, mouse and rat Phospho-Mek1 (Ser217/221).
    Compatible Sample Types Tissue Lysates, Cell Lysates
    Solid Support 96-well Microplate
    Method Of Detection Colorimetric
    Design Principle Sandwich-based
    Research Area Post-Translational Modifications, Phosphorylation, MAPK Signaling
    Estimated Lead Time 1-2 business days
    Shipping Type Blue ice
    Storage -20°C

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    Product Features

    • Rapidly measure phosphorylated protein in lysates
    • Screen numerous different cell lysates without performing a Western Blot analysis
    • Minimal hands-on time, convenient, and non-radioactive material

    Application Notes

    Kit Components
    • Pre-Coated 96-well Strip Microplate
    • Wash Buffer
    • Anti-Phospho Antibody
    • HRP-Conjugated Secondary Antibody
    • Assay Diluent
    • TMB One-Step Substrate
    • Stop Solution
    • Lysis Buffer
    • Positive Control Sample
    Other Materials Required
    • Distilled or deionized water
    • 100 ml and 1 liter graduated cylinders
    • Tubes to prepare sample dilutions
    • Protease and Phosphatase inhibitors
    • Precision pipettes to deliver 2 µl to 1 ml volumes
    • Adjustable 1-25 ml pipettes for reagent preparation
    • Benchtop rocker or shaker
    • Microplate reader capable of measuring absorbance at 450 nm
    Protocol Outline
    1. Prepare all reagents and samples as instructed in the manual.
    2. Add 100 µl of sample or positive control to each well.
    3. Incubate 2.5 h at RT or O/N at 4 °C.
    4. Add 100 µl of prepared primary antibody to each well.
    5. Incubate 1 h at RT.
    6. Add 100 µl of prepared 1X HRP-Streptavidin to each well.
    7. Incubate 1 h at RT.
    8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
    9. Incubate 30 min at RT.
    10. Add 50 µl of Stop Solution to each well.
    11. Read at 450 nm immediately.

    Storage/Stability

    Upon receipt, the kit should be stored at -20 °C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), HRP-Streptavidin (Item G), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4 °C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge and store at -20 °C. Reconstituted Positive Control (Item K) should be stored at -70 °C.

    Citations

    Tunctan, Bahar, et al. "5, 14-HEDGE, a 20-HETE mimetic, reverses hypotension and improves survival in a rodent model of septic shock: Contribution of soluble epoxide hydrolase, CYP2C23, MEK1/ERK1/2/IKK?/I?B-?/NF-?B pathway, and proinflammatory cytokine formation." Prostaglandins & other lipid mediators 102 (2013): 31-41.
    Species:
    Mouse
    Sample Type:
    Tissue Lysate (kidneys from Endotoxic shock )
    View Publication
    Tunctan, Bahar, et al. "5, 14-HEDGE, a 20-HETE mimetic, reverses hypotension and improves survival in a rodent model of septic shock: Contribution of soluble epoxide hydrolase, CYP2C23, MEK1/ERK1/2/IKK?/I?B-?/NF-?B pathway, and proinflammatory cytokine formation." Prostaglandins & other lipid mediators 102 (2013): 31-41.
    Species:
    Rat
    Sample Type:
    Tissue Lysate (kidneys from Endotoxic shock )
    View Publication

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