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Intermediate filament proteins /
Intermediate Filament Proteins, the latest volume in the Methods in Enzymology series covers all the intermediate filaments in vertebrates and invertebrates, providing a unique understanding of the multiple different tissue-specific intermediate filaments. This volume also covers the latest methods...
| Clasificación: | Libro Electrónico |
|---|---|
| Otros Autores: | , |
| Formato: | Electrónico eBook |
| Idioma: | Inglés |
| Publicado: |
Waltham, MA :
Academic Press, an imprint of Elsevier,
2016.
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| Colección: | Methods in enzymology ;
v. 568. |
| Temas: | |
| Acceso en línea: | Texto completo Texto completo |
Tabla de Contenidos:
- Front Cover; Intermediate Filament Proteins; Copyright; Dedication; Contents; Contributors; Preface; References; Acknowledgments; Part I: General Methods to Study IF Proteins; Chapter One: How to Study Intermediate Filaments in Atomic Detail; 1. Introductory Remarks; 2. Bioinformatics; 3. X-ray Crystallography; 3.1. Design of IF Protein Fragments for Crystallization; 3.2. Protein Expression; 3.3. Purification; 3.4. Crystallization; 3.5. X-ray Data Collection; 3.6. Phasing by Molecular Replacement; 3.7. Experimental Phasing Using Heavy Atoms; 3.8. Structure Refinement and Validation.
- 3.9. Limitations and Pitfalls3.10. Obtained Structures and Impact; 4. Electron Paramagnetic Resonance with Site-Directed Spin Labeling; 4.1. Principle; 4.2. Sample Preparation and Measurements; 4.3. Data Interpretation and Impact; 4.4. Limitations and Outlook; 5. Concluding Remarks; Acknowledgments; References; Chapter Two: Mechanical Properties of Intermediate Filament Proteins; 1. Introduction; 2. Viscoelasticity of Purified IFs In Vitro; 3. IFs and the Mechanical Properties of Cells; 3.1. Type III IF: Vimentin and Desmin; 3.1.1. Knockout Models of Vimentin.
- 3.1.2. Drugs and Proteins Disrupting Vimentin3.1.3. Mutated Desmin to Disrupt Vimentin; 3.1.4. Mutated Desmin to Disrupt Desmin; 3.2. Keratins; 3.3. Neurofilaments; 4. Conclusion; Acknowledgments; References; Chapter Three: Multidimensional Monitoring of Keratin Intermediate Filaments in Cultured Cells and Tissues; 1. Introduction; 2. 3D Imaging of Keratin Intermediate Filaments in Cultured Cells; 2.1. Microscopes; 2.2. Preparation of Imaging Medium; 2.3. Surface Coating; 2.4. Preparation of Cells for Imaging; 2.5. Image Acquisition; 2.6. Keratin Network Normalization.
- 2.7. Measuring Keratin Movement2.8. Keratin Bulk Flow Analysis; 2.9. Keratin Turnover Measured by FRAP; 3. 3D Imaging of Keratin Intermediate Filaments in Murine Preimplantation Embryos; 3.1. Embryo Collection and Cultivation; 3.2. Imaging of Preimplantation Embryos; 3.3. Image Processing and Analysis; 4. Outlook; Acknowledgments; References; Chapter Four: Phospho-Specific Antibody Probes of Intermediate Filament Proteins; 1. Introduction; 2. Production of Site- and Phosphorylation State-Specific Antibodies; 2.1. Designing a Synthetic Phosphopeptide.
- 2.2. Conjugation of a Phosphopeptide to a Carrier Protein3. Characterization of Site- and Phosphorylation State-Specific Antibodies; 3.1. ELISA; 3.2. Immunoblotting; 4. Immunocytochemistry; 4.1. Overview for Immunostaining Methods; 4.2. Vimentin Phosphorylation in Mitosis; 4.3. Vimentin Phosphorylation in Signal Transduction; 5. Other Applications; 6. Conclusions; Acknowledgments; References; Chapter Five: Assays for Posttranslational Modifications of Intermediate Filament Proteins; 1. Introduction; 1.1. Posttranslational Modifications of Intermediate Filament Proteins.