Nuclear pore complexes and nucleocytoplasmic transport: methods
Front Cover; Nuclear Pore Complexes and Nucleocytoplasmic Transport -- Methods; Copyright; Contents; Contributors; Preface; Chapter 1: Fifty Years of Nuclear Pores and Nucleocytoplasmic Transport Studies: Multiple Tools Revealing Complex Rules; Introduction; 1.1. The NPCs: A Modular Macromolecular A...
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| Format: | Book/Monograph |
| Language: | English |
| Published: |
San Diego, CA
Academic Press
2014
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| Edition: | First edition |
| Series: | Methods in cell biology
122 |
| In: |
Methods in cell biology (122)
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| Volumes / Articles: | Show Volumes / Articles. |
| Subjects: | |
| Online Access: | Verlag, Volltext: http://www.sciencedirect.com/science/book/9780124171602 Verlag, Volltext: http://www.sciencedirect.com/science/bookseries/0091679X/122 Aggregator, Inhaltsverzeichnis: https://www.gbv.de/dms/bowker/toc/9780124171602.pdf 
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| Author Notes: | ed. by Valérie Doye |
| Summary: | Front Cover; Nuclear Pore Complexes and Nucleocytoplasmic Transport -- Methods; Copyright; Contents; Contributors; Preface; Chapter 1: Fifty Years of Nuclear Pores and Nucleocytoplasmic Transport Studies: Multiple Tools Revealing Complex Rules; Introduction; 1.1. The NPCs: A Modular Macromolecular Assembly; 1.1.1. Toward a refined view of the NPC structure; 1.1.2. Nucleoporins: The building blocks of NPCs; 1.1.3. Integrating the Nups into the 3D architecture of the NPC; 1.1.3.1. Nup localization; 1.1.3.2. Interactions among Nups; 1.1.3.3. Nup stoichiometry Chapter 2: Imaging Metazoan Nuclear Pore Complexes by Field Emission Scanning Electron MicroscopyIntroduction; Prehistoric landmarks (before EM): From cells to the nuclear envelope; The EM era: From the nuclear envelope to nuclear pore complexes; High resolution SEM: Direct surface imaging of NPCs; 2.1. Rationale; 2.1.1. Purpose; 2.1.2. An overview of sample preparation; 2.1.2.1. Fixation; 2.1.2.2. Dehydration and critical point drying; 2.1.2.3. Sputter coating; 2.2. Materials; 2.2.1. Equipment; 2.2.2. Materials; 2.2.3. Reagents; 2.2.4. Buffers; 2.3. Anchored Nuclei 2.3.1. Coating silicon chips2.3.2. Chromatin decondensation and attachment; 2.3.3. In vitro assembly reaction; 2.4. Mammalian Cell Nuclei; 2.5. Immunogold Labeling; 2.6. Sample Preparation for FESEM; 2.6.1. Fixation, dehydration, and CPD; 2.6.2. Sputter coating and FESEM analysis; Acknowledgments; References; Chapter 3: Imaging Yeast NPCs: From Classical Electron Microscopy to Immuno-SEM; Introduction; 3.1. Conventional TEM; 3.1.1. Materials; 3.1.1.1. Equipment; 3.1.1.2. Spheroplast preparation; 3.1.1.3. Fixation, dehydration, contrasting, and embedding; 3.1.1.4. Sectioning and poststaining Volume 122 of Methods in Cell Biology describes modern tools and techniques used to study nuclear pore complexes and nucleocytoplasmic transport in diverse eukaryotic model systems (including mammalian cells, Xenopus, C. elegans, yeast). The volume enables investigators to analyze nuclear pore complex structure, assembly, and dynamics; to evaluate protein and RNA trafficking through the nuclear envelope; and to design in vivo or in vitro assays appropriate to their research needs. Beyond the study of nuclear pores and transport as such, these protocols will also |
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| Item Description: | Description based upon print version of record. - 3.1.2. Procedure |
| Physical Description: | Online Resource |
| ISBN: | 9780124171787 0124171788 |