Septins provides established septin and molecular and developmental biologists and researchers new to the field with proven, state-of-art techniques and relevant historical background and theory to aid efficient design and effective implementation of experimental methodologies. Topics include the purification of septin proteins from diverse systems, their visualization in live cells, and their analysis by a variety of cutting-edge microscopy approaches.
1. Cover image - p. i 2. Title page - p. i 3. Table of Contents - p. iii 4. Series Editors - p. ii 5. Copyright - p. iv 6. Contributors - p. xi 7. Preface - p. xv 8. Chapter 1. The nonopisthokont septins: How many there are, how little we know about them, and how we might learn more - p. 1 9. Chapter 2. Preparing recombinant yeast septins and their analysis by electron microscopy - p. 21 10. Chapter 3. A FRET-based method for monitoring septin polymerization and binding of septin-associated proteins - p. 35 11. Chapter 4. In vitro reconstitution of septin assemblies on supported lipid bilayers - p. 57 12. Chapter 5. Visualization of in vivo septin ultrastructures by platinum replica electron microscopy - p. 73 13. Chapter 6. Assays for genetic dissection of septin filament assembly in yeast, from de novo folding through polymerization - p. 99 14. Chapter 7. Investigation of septins using infection by bacterial pathogens - p. 117 15. Chapter 8. In vivo analysis of septin heteropolymer rods and higher-order structures in filamentous fungi - p. 135 16. Chapter 9. Live cell imaging of septin dynamics in Ustilago maydis - p. 143 17. Chapter 10. Ashbya gossypii as a model system to study septin organization by single-molecule localization microscopy - p. 161 18. Chapter 11. Visualizing septins in early Drosophila embryos - p. 183 19. Chapter 12. Purification of recombinant human and Drosophila septin hexamers for TIRF assays of actin–septin filament assembly - p. 199 20. Chapter 13. Investigation of septin biology in vivo using zebrafish - p. 221 21. Chapter 14. Fluorescence microscopy of actin- and microtubule-associated septins in mammalian cells - p. 243 22. Chapter 15. Immunofluorescent staining of septins in primary cilia - p. 269 23. Chapter 16. Methods for immunoblot detection and electron microscopic localization of septin subunits in mammalian nervous systems - p. 285 24. Chapter 17. Visualizing septin and cell dynamics in mammalian brain slices - p. 295 25. Chapter 18. Small molecule perturbations of septins - p. 311 26. Chapter 19. Septin crystallization for structural analysis - p. 321 27. Index - p. 339 28. Volumes in Series - p. 353
Septins provides established septin and molecular and developmental biologists and researchers new to the field with proven, state-of-art techniques and relevant historical background and theory to aid efficient design and effective implementation of experimental methodologies. Topics include the purification of septin proteins from diverse systems, their visualization in live cells, and their analysis by a variety of cutting-edge microscopy approaches.
1. Cover image - p. i 2. Title page - p. i 3. Table of Contents - p. iii 4. Series Editors - p. ii 5. Copyright - p. iv 6. Contributors - p. xi 7. Preface - p. xv 8. Chapter 1. The nonopisthokont septins: How many there are, how little we know about them, and how we might learn more - p. 1 9. Chapter 2. Preparing recombinant yeast septins and their analysis by electron microscopy - p. 21 10. Chapter 3. A FRET-based method for monitoring septin polymerization and binding of septin-associated proteins - p. 35 11. Chapter 4. In vitro reconstitution of septin assemblies on supported lipid bilayers - p. 57 12. Chapter 5. Visualization of in vivo septin ultrastructures by platinum replica electron microscopy - p. 73 13. Chapter 6. Assays for genetic dissection of septin filament assembly in yeast, from de novo folding through polymerization - p. 99 14. Chapter 7. Investigation of septins using infection by bacterial pathogens - p. 117 15. Chapter 8. In vivo analysis of septin heteropolymer rods and higher-order structures in filamentous fungi - p. 135 16. Chapter 9. Live cell imaging of septin dynamics in Ustilago maydis - p. 143 17. Chapter 10. Ashbya gossypii as a model system to study septin organization by single-molecule localization microscopy - p. 161 18. Chapter 11. Visualizing septins in early Drosophila embryos - p. 183 19. Chapter 12. Purification of recombinant human and Drosophila septin hexamers for TIRF assays of actin–septin filament assembly - p. 199 20. Chapter 13. Investigation of septin biology in vivo using zebrafish - p. 221 21. Chapter 14. Fluorescence microscopy of actin- and microtubule-associated septins in mammalian cells - p. 243 22. Chapter 15. Immunofluorescent staining of septins in primary cilia - p. 269 23. Chapter 16. Methods for immunoblot detection and electron microscopic localization of septin subunits in mammalian nervous systems - p. 285 24. Chapter 17. Visualizing septin and cell dynamics in mammalian brain slices - p. 295 25. Chapter 18. Small molecule perturbations of septins - p. 311 26. Chapter 19. Septin crystallization for structural analysis - p. 321 27. Index - p. 339 28. Volumes in Series - p. 353
