Intermediate Filament Cytoskeleton

Front Cover
Gulf Professional Publishing, 2004 - Medical - 886 pages
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Intermediate filaments are a large family of proteins that are the cytoskeletal elements involved in a number of skin, liver, neuromuscular, cardiac, eye and hair diseases. Intermediate filament genes are regulated in a tissue-and cell type-specific manner and their polymerized protein products protects the cells and tissue they are part of against a variety of mechanical and nonmechanical stresses. This book provides a comprehensive resource of methodology essentials, describing a variety of essential tools and assays for studying intermediate filaments. The book provides user-friendly advice and protocols covering all aspects of intermediate filaments including protein isolation and structure, protein and gene regulation, relationship to disease and apoptosis, and associated proteins. Both mammalian and non-mammalian systems and animal models are covered, making this book a must-have for any investigator wishing to study IF genes or their protein products.

* Covers intermediate filaments from crystallography, protein chemistry, cell and molecular biology, microrheology, gene regulation, to animal models and human disease
* Practical and user-friendly with detailed "how-to-protocols and "tricks of the trade"
* Includes detailed tables of useful reagents, vendors and web links
 

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Contents

Isolation Characterization and In Vitro Assembly of Intermediate Filaments
3
Intermediate Filament Protein Structure Determination
25
Intracellular Microrheology as a Tool for the Measurement of the Local
45
Developing Mouse Models to Study Intermediate Filament Function
65
Uncovering the Roles of Intermediate Filaments in Apoptosis
95
Molecular Genetics Methods for Human Intermediate Filament Diseases
131
Intermediate Filaments and Multiparameter Flow Cytometry
163
Procedures
191
Muscle Intermediate Filament Proteins
519
Pearls and Pitfalls
547
Concluding Remarks
548
References
549
SIMethionine Metabolic Labeling to Study Axonal Transport of Neuronal Intermediate Filament Proteins In Vivo Stéphanie Millecamps and JeanPier...
555
Materials and Instrumentation
560
Procedures
561
Comments
565

Comments
196
Pearls and Pitfalls
198
References
199
Intermediate Filament Protein Inclusions Kurt Zatloukal Conny Stumptner Andrea Fuchsbichler Elke Janig and Helmut Denk
205
Introduction
206
Materials and Instrumentation
216
Procedures
218
Pitfalls
224
References
225
Stress Models for the Study of Intermediate Filament Function E Birgitte Lane and Milos Pekny
229
Introduction
230
Materials and Instrumentation
249
Procedures
251
Pitfalls
256
DiscussionConcluding Remarks
258
General Methods to Study Intermediate Filament Proteins and Gene Regulation
265
Regulation of Intermediate Filament Gene Expression Satrajit Sinha I Introduction
267
Materials
277
Procedures
278
Pitfalls
292
Discussion and Concluding Remarks
293
FluorescenceBased Methods for Studying Intermediate Filaments
297
Imaging of Keratin Dynamics during the Cell Cycle and in Response
321
Approaches to Study Phosphorylation of Intermediate Filament Proteins Using
353
Approaches to Study Posttranslational Regulation of Intermediate Filament Proteins
373
Methods to Study Specific Intermediate Filament
411
An Ideal Model System to Study Keratin Genes and Proteins
453
Studying Simple Epithelial Keratins in Cells and Tissues
489
Pitfalls
566
Lamins Georg Krohne
573
Introduction
574
Materials
576
Purification of Lamins Produced in Bacteria
579
Production of Lamins in Insect Cells Using the Baculovirus System
586
Selection of Cultured Cells and Tissues for Lamin Enrichment
588
Enrichment of Lamins from Cultured Cells Tested for Xenopus A6 Cells and HeLa Cells
591
Immunoprecipitation of Lamin from Cell Extracts
593
References
594
The Intermediate Filament Systems in the Eye Lens Ming Der Perng Aileen Sandilands Jer Kuszak Ralf Dahm Alfred Wegener Alan R Prescott and R...
597
Introduction
598
Materials and Instrumentation
603
Procedures
607
Pearls and Pitfalls
620
References
621
Methods to Study Specific Intermediate Filament
625
Performing Functional Studies of Xenopus laevis Intermediate Filament Proteins
673
Intermediate Filaments in Caenorhabditis elegans
703
Live Imaging of C elegans 71 4
714
Methods to Study Intermediate
719
In Vitro Methods for Investigating DesmoplakinIntermediate Filament
757
Studying Cytolinker Proteins
787
Keratin Bundling Proteins
817
Index
861
Volumes in Series
879
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