Umfang:
Online-Ressource (XXV, 791 Seiten)
,
Illustrationen
Ausgabe:
First edition
Ausgabe:
Online-Ausg.
ISBN:
012803470X
,
0128034912
,
9780128034910
Serie:
Methods in Enzymology volume 568
Inhalt:
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 that are currently being used to study intermediate filament protein function and dynamics. It will be an important companion for any experimentalist interesting in studying this protein family in their cell or organism model system
Inhalt:
Intermediate Filament Proteins, the latest volume in the Methods in Enzymologyseries 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 that are currently being used to study intermediate filament protein function and dynamics. It will be an important companion for any experimentalist interesting in studying this protein family in their cell or organism model system.Focuses on intermediate filaments, including the latest information Provides an up-to-date understanding of the fieldContains contributions from the major scientists working and publishing in the field
Anmerkung:
Description based upon print version of record
,
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
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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
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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
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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
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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
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1.2. Available Tools and Major Limitations for the Study of IF Protein PTMs
Weitere Ausg.:
ISBN 012803470X
Weitere Ausg.:
ISBN 9780128034705
Weitere Ausg.:
Print version Intermediate Filament Proteins
Sprache:
Englisch
Fachgebiete:
Biologie
Schlagwort(e):
Intermediärfilament
;
Proteine
;
Electronic books
;
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