Book Volume 1487 2017

ERK Signaling

Methods and Protocols

Editors:

ISBN: 978-1-4939-6422-2 (Print) 978-1-4939-6424-6 (Online)

Table of contents (29 protocols)

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  1. Front Matter

    Pages i-xii

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    Protocol

    Pages 1-21

    How Genetics Has Helped Piece Together the MAPK Signaling Pathway

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    Protocol

    Pages 23-33

    In Vitro Enzyme Kinetics Analysis of EGFR

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    Protocol

    Pages 35-52

    High-Throughput Analysis of Mammalian Receptor Tyrosine Kinase Activation in Yeast Cells

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    Protocol

    Pages 53-63

    Structural Studies of ERK2 Protein Complexes

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    Protocol

    Pages 65-88

    Isolation and Characterization of Intrinsically Active (MEK-Independent) Mutants of Mpk1/Erk

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    Protocol

    Pages 89-98

    Assaying Activation and Subcellular Localization of ERK in Cells and Tissues

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    Protocol

    Pages 99-111

    Detection and Functional Analysis of SUMO-Modified MEK

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    Protocol

    Pages 113-126

    Single-Step Affinity Purification of ERK Signaling Complexes Using the Streptavidin-Binding Peptide (SBP) Tag

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    Protocol

    Pages 127-135

    High-Throughput In Vitro Identification of Direct MAPK/Erk Substrates

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    Protocol

    Pages 137-149

    Global Identification of ERK Substrates by Phosphoproteomics Based on IMAC and 2D-DIGE

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    Protocol

    Pages 151-162

    Analysis of Ras/ERK Compartmentalization by Subcellular Fractionation

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    Pages 163-174

    Cell-Based Assays to Study ERK Pathway/Caveolin1 Interactions

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    Protocol

    Pages 175-194

    The Nuclear Translocation of ERK

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    Protocol

    Pages 195-201

    Visualization of RAS/MAPK Signaling In Situ by the Proximity Ligation Assay (PLA)

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    Protocol

    Pages 203-221

    Measuring ERK Activity Dynamics in Single Living Cells Using FRET Biosensors

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    Protocol

    Pages 223-234

    Quantifying Tensile Force and ERK Phosphorylation on Actin Stress Fibers

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    Protocol

    Pages 235-241

    Co-culture Activation of MAP Kinase in Drosophila S2 Cells

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    Protocol

    Pages 243-253

    Isolation of Mouse Embryonic Stem Cell Lines in the Study of ERK1/2 MAP Kinase Signaling

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    Protocol

    Pages 255-267

    3D Organotypic Culture Model to Study Components of ERK Signaling

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    Protocol

    Pages 269-276

    Genetic Validation of Cell Proliferation via Ras-Independent Activation of the Raf/Mek/Erk Pathway

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