JAK-STAT Signalling

Methods and Protocols

Editors:

ISBN: 978-1-62703-241-4 (Print) 978-1-62703-242-1 (Online)

Table of contents (21 protocols)

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  1. Front Matter

    Pages i-xi

  2. JAK Protein Tyrosine Kinases

    1. Front Matter

      Pages 1-1

    2. No Access

      Book Protocol

      Pages 3-20

      Analysis of Janus Tyrosine Kinase Phosphorylation and Activation

    3. No Access

      Book Protocol

      Pages 21-38

      Co-immunoprecipitation Protocol to Investigate Cytokine Receptor-Associated Proteins, e.g., Janus Kinases or Other Associated Signaling Proteins

    4. No Access

      Book Protocol

      Pages 39-55

      In Vitro JAK Kinase Activity and Inhibition Assays

    5. No Access

      Book Protocol

      Pages 57-67

      Quantitative Analysis of JAK Binding Using Isothermal Titration Calorimetry and Surface Plasmon Resonance

    6. No Access

      Book Protocol

      Pages 69-80

      Determination of Protein Turnover Rates in the JAK/STAT Pathway Using a Radioactive Pulse-Chase Approach

    7. No Access

      Book Protocol

      Pages 81-97

      Designing RNAi Screens to Identify JAK/STAT Pathway Components

    8. No Access

      Book Protocol

      Pages 99-113

      The Use of JAK-Specific Inhibitors as Chemical Biology Tools

    9. No Access

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      Pages 115-131

      Methods for Detecting Mutations in the Human JAK2 Gene

  3. Signal Transducers and Activators of Transcription (STAT)s

    1. Front Matter

      Pages 133-133

    2. No Access

      Book Protocol

      Pages 135-145

      Detection of Activated STAT Proteins

    3. No Access

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      Pages 147-159

      Detection of Activated STAT Species Using Electrophoretic Mobility Shift Assay (EMSA) and Potential Pitfalls Arising from the Use of Detergents

    4. No Access

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      Pages 161-165

      Flow Cytometric Analysis of STAT Phosphorylation

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      Pages 167-178

      Acetylation of Endogenous STAT Proteins

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      Pages 179-188

      Detection and Cellular Localization of Phospho-STAT2 in the Central Nervous System by Immunohistochemical Staining

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      Pages 189-202

      Nuclear Trafficking of STAT Proteins Visualized by Live Cell Imaging

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      Pages 203-224

      Characterization of STAT Self-Association by Analytical Ultracentrifugation

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      Pages 225-232

      Constitutively Active STAT5 Constructs

  4. Suppressor of Cytokine Signalling (SOCS) Proteins

    1. Front Matter

      Pages 233-233

    2. No Access

      Book Protocol

      Pages 235-248

      Analysis of Suppressor of Cytokine Signalling (SOCS) Gene Expression by Real-Time Quantitative PCR

    3. No Access

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      Pages 249-259

      Detection of Endogenous SOCS1 and SOCS3 Proteins by Immunoprecipitation and Western Blot Analysis

    4. No Access

      Book Protocol

      Pages 261-271

      In Vitro Ubiquitination of Cytokine Signaling Components

  5. Structural Analysis of the JAK/STAT Pathway

    1. Front Matter

      Pages 273-273

    2. No Access

      Book Protocol

      Pages 275-300

      Production and Crystallization of Recombinant JAK Proteins

  6. Back Matter

    Pages 319-320

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