Book Volume 1557 2017

CD95

Methods and Protocols

Editors:

ISBN: 978-1-4939-6778-0 (Print) 978-1-4939-6780-3 (Online)

Table of contents (22 protocols)

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  1. Front Matter

    Pages i-xi

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    Protocol

    Pages 1-10

    Production of the Non-apoptotic Metalloprotease-Cleaved CD95L and Its Cytotoxic Recombinant Counterpart Designed Ig-CD95L

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    Protocol

    Pages 11-18

    CD95 Stimulation with CD95L and DISC Analysis

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    Protocol

    Pages 19-31

    Immunoprecipitation of Death Inducing Signaling Complex by Caspase-8

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    Protocol

    Pages 33-40

    In Vitro Evaluation of the Apoptosis Function in Human Activated T Cells

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    Protocol

    Pages 41-48

    Proximity Ligation Assay (PLA) to Evaluate DISC and MISC Composition

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    Protocol

    Pages 49-62

    Fluorometric Methods for Detection of Mitochondrial Membrane Depolarization Induced by CD95 Activation

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    Protocol

    Pages 63-77

    Generation and Application of Bioluminescent CD95 Ligand Fusion Proteins

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    Protocol

    Pages 79-93

    CD95-Mediated Calcium Signaling

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    Protocol

    Pages 95-102

    CD95-Mediated Proton Regulation

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    Protocol

    Pages 103-110

    Study of the CD95-Mediated Non-apoptotic Signaling Pathway: PI3K

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    Protocol

    Pages 111-115

    Organelle Separation and Cell Signaling

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    Protocol

    Pages 117-123

    Boyden Chamber Assay to Study of Cell Migration Induced by Metalloprotease Cleaved-CD95L

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    Protocol

    Pages 125-138

    Isolation of Lipid Rafts Through Discontinuous Sucrose Gradient Centrifugation and Fas/CD95 Death Receptor Localization in Raft Fractions

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    Protocol

    Pages 139-151

    Quantifying CD95/cl-CD95L Implications in Cell Mechanics and Membrane Tension by Atomic Force Microscopy Based Force Measurements

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    Protocol

    Pages 153-171

    Sketching of CD95 Oligomers by In Silico Investigations

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    Protocol

    Pages 173-188

    Site-Specific Detection of Tyrosine Phosphorylated CD95 Following Protein Separation by Conventional and Phospho-Protein Affinity SDS-PAGE

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    Protocol

    Pages 189-198

    Detection of S-Acylated CD95 by Acyl-Biotin Exchange

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    Protocol

    Pages 199-206

    Exploration of Fas S-Nitrosylation by the Biotin Switch Assay

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    Protocol

    Pages 207-212

    Method to Measure Sphingomyelin Synthase Activity Changes in Response to CD95L

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    Protocol

    Pages 213-217

    Liquid Chromatography–High Resolution Mass Spectrometry Method to Study Sphingolipid Metabolism Changes in Response to CD95L

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