The Cytoskeleton, Part A
Cytoskeleton Proteins, Isolation and Characterization
Academic Press
Published on 4. August 1982
444 pages
978-0-08-085922-4 (ISBN)
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The Cytoskeleton, Part A
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05/1982
Academic Press
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Content
- Front Cover
- Methods in Cell Biology, Volume 24
- Copyright Page
- Contents
- Contributors
- Preface
- Chapter 1. The Cytoskeleton: A Perspective
- I. Microtubules
- II. Microfilaments
- III. Intermediate Filaments
- References
- Chapter 2. Purification and Chemical Properties of Brain Tubulin
- I. Introduction
- II. Purification of Brain Tubulin
- III. Chemical Properties of Brain Tubulin
- IV. Posttranslational Modification
- V. Conclusion
- References
- Chapter 3. Assembly -Disassembly Purification and Characterization of Microtubule Protein without Glycerol
- I. Introduction
- II. General Remarks on the Isolation of Microtubule Protein
- III. Purification of Microtubule Protein by Cycles of Assembly and Disassembly
- IV. Purification of Tubulin and MAPs from Microtubule Protein
- V. Perspectives on the Role of MAPs in Microtubule Assembly and Function
- References
- Chapter 4. A Brain Microtubule Protein Preparation Depleted of Mitochondrial and Synaptosomal Components
- I. Introduction
- II. Experimental Procedures
- III. Molecular Characteristics of Purified Microtubule Protein
- IV. Concluding Remarks
- References
- Chapter 5. Purification and Reassembly of Tubulin from Outer Doublet Microtubules
- I. Introduction
- II. Methodologies
- III. Reassembly Properties of Outer Doublet Tubulin
- IV. Properties of Microtubules Reassembled from Outer Doublet Tubulin
- V. Conclusions
- References
- Chapter 6. Production of Antisera and Radioimmunoassays for Tubulin
- I. Introduction
- II. Methods
- III. Results
- IV. Discussion
- References
- Chapter 7. Immunofluorescence and Immunocytochemical Procedures with Affinity Purified Antibodies: Tubulin-Containing Structures
- I. Introduction
- II. Antibody Production, Purification, and Characterization
- III. Direct versus Indirect Immunofluorescence Microscopy
- IV. Indirect Immunofluorescence Technique
- V. Double Immunofluorescence Microscopy
- VI. Use of Glutaraldehyde in Immunofluorescence Microscopy
- VII. Comparison of the Same Specimen in Immunofluorescence Microscopy and in Electron Microscopy
- VIII. Limit of Resolution of Fluorescence Microscopy
- IX. Stereo Immunofluorescence Microscopy
- X. Peroxidase Immunoferritin and Gold Labeling
- References
- Chapter 8. An Automated Method for Defining Microtubule Length Distributions
- I. Introduction
- II. Experimental Procedures
- III. Discussion
- References
- Appendix: Length Distribution Program for the HP 9825A
- Chapter 9. Measurement of Steady-State Tubulin Flux
- I. Introduction
- II. Use of GTP as a Probe of the Equilibrium State
- III. A Simple Approach to Measurement of Treadmilling Rates
- IV. Other Methods for Determination of Rates, Rate Constants, and Efficiency of Treadmilling
- References
- Chapter 10. A Rapid Filtration Assay for Analysis of Microtubule Assembly, Disassembly, and Steady-State Tubulin Flux
- I. Introduction
- II. Rapid Filtration Assay: Details of the Procedure
- III. Characterization of the Assay System
- IV. Concluding Remarks
- References
- Chapter 11. A Direct Method for Analyzing the Polymerization Kinetics at the Two Ends of a Microtubule
- I. Introduction
- II. Materials and Methods
- III. Results and Discussion
- IV. Perspective
- References
- Chapter 12. Dynein Decoration of Microtubules-Determination of Polarity
- I. Introduction
- II. Methods
- III. Analysis of Dynein Binding to Microtubules
- IV. Analysis of Microtubule Polarity Using Bound Flagellar Dynein
- V. Perspectives
- References
- Chapter 13. Microtubule Polarity Determination Based on Conditions for Tubulin Assembly in Vitro
- I. Introduction
- II. Overview of the Method
- III. Conditions for Microtubule Assembly in Vitro That Promote Hook Formation
- IV. Formation of Hooks on Cytoplasmic Microtubules
- V. Electron Microscopy
- VI. Interpretation of Hook Handedness
- References
- Chapter 14. The Use of Tannic Acid in Microtubule Research
- I. Introduction
- II. Microtubule Substructure-Brief Review
- Ill. Use of Tannic Acid in Microtubule Research
- IV. Use of Tannic Acid in Other Ultrastructural Studies
- V. Conclusion
- References
- Chapter 15. The Cyclic Tyrosination/Detyrosination of Alpha Tubulin
- I. Introduction
- II. Review
- III. Methods
- IV. Perspectives
- References
- Chapter 16. Tubulin -Tyrosine Ligase from Brain
- I. Introduction
- II. Assay Method
- III. Purification Procedure
- IV. Properties
- References
- Chapter 17. Preparation of Brain Tyrosinotubulin Carboxypeptidase
- I. Introduction
- II. Assay
- Ill. Purification of Brain Tyrosinotubulin Carboxypeptidase
- IV. Remarks
- References
- Chapter 18. Purifcution of Muscle Actin
- I. Introduction
- II. Acetone Powder Preparation
- Ill. Actin Isolation
- IV. Analysis of Purity of Final Product
- V. Sources of Contamination
- VI. Further Purification of Actin
- VII. Concluding Remarks
- References
- Chapter 19. Directional Growth of Actin Off Polylysine-Coated Beads
- I. Introduction-An Overview of the Actin Assembly Reaction
- II. Preparation of Polylysine-Coated Polystyrene Beads
- III. Nature of the Actin-Bead Interaction
- IV. Summary of the Usefulness of Polylysine Beads as a Tool for Studying Actin
- References
- Chapter 20. A Falling Ball Apparatus to Measure Filament Cross-Linking
- I. Introduction
- II. Theory
- III. Design and Operation of the Falling Ball Apparatus
- IV. Testing and Calibration of the Falling Ball Apparatus
- V. Application of the Falling Ball Viscometer to Non-Newtonian Fluids
- VI. Practical Applications of the Failing Ball Apparatus
- VII. Conclusion: Evaluation and Comparison with Other Assays
- References
- Chapter 21. Antibody Production and Immunofluorescent Characterization of Actin and Contractile Proteins
- I. Introduction
- II. Methods
- III. Immunological Characterization of Antiactin Antibodies
- IV. Autoimmune Antibodies to Actin
- V. Immunofluorescent Distribution of Actin in Cells Grown in Tissue Culture
- References
- Chapter 22. Myosin Purification and Characterization
- I. Introduction
- II. Myosin Assays
- III. Myosin Extraction from Cells
- IV. Fractional Precipitation Methods
- V. Chromatography of Myosin
- VI. Concentration
- VII. Storage
- VIII. Detailed Methods to Purify Specific Myosins
- IX. Perspectives
- References
- Chapter 23. Preparation and Purification of Dynein
- I. Introduction
- II. Preparation of Latent Activity Dynein-1
- Ill. Further Purification of Dynein
- IV. Selected Properties of LAD-1 and Its Subunits
- V. Preparation and Properties of Dynein ATPases from Other Sources
- VI. Summary
- References
- Chapter 24. Isolation and Characterization of Intermediate Filaments
- I. Introduction
- II. IF of Specific Types of Cells
- III. Chemical and Structural Features of IF
- References
- Index
- Contents of Recent Volumes
