The Nuclear Envelope: Methods and Protocols

The Nuclear Envelope: Methods and Protocols

Paperback(Softcover reprint of the original 1st ed. 2016)

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This volume provides a wide range of protocols used in studying the nuclear envelope, with special attention to the experimental adjustments that may be required to successfully investigate this complex organelle in cells from various organisms. The Nuclear Envelope:
Methods and Protocols
is divided into five sections: Part I – Nuclear Envelope
Isolation; Part II – Nuclear Envelope Protein Interactions, Localization, and
Dynamics; Part III – Nuclear Envelope Interactions with the Cytoskeleton; Part
IV – Nuclear Envelope-Chromatin Interactions; and Part V – Nucleo-Cytoplasmic
Transport. Many of the modifications discussed in this book have only been circulated within laboratories that have conducted research in this field for many years. Written in the highly successful Methods in Molecular Biology
series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting edge and thorough, The Nuclear Envelope: Methods and Protocols is a timely resource for researchers who have joined this dynamic and rapidly growing field.

Product Details

ISBN-13: 9781493980680
Publisher: Springer New York
Publication date: 05/26/2018
Series: Methods in Molecular Biology , #1411
Edition description: Softcover reprint of the original 1st ed. 2016
Pages: 523
Product dimensions: 7.01(w) x 10.00(h) x (d)

Table of Contents

Part I Nuclear Envelope Isolation

  1. Isolation,
    Proteomic Analysis and Microscopy Confirmation of the Liver Nuclear Envelope

    Nadia Korfali, Laurence
    Florens, and Eric C. Schirmer

  2. Exploring the Protein Composition of the Plant Nuclear Envelope

    Xiao Zhou,
    Kentaro Tamura, Katja Graumann, and Iris Meier

  3. High Efficiency
    Isolation of Nuclear Envelope Protein Complexes from Trypanosomes

    Samson Obado, Mark C.
    Field, Brian T. Chait, and Michael P. Rout

II Nuclear Envelope Protein Interactions, Localization, and Dynamics

4. Super Resolution Microscopy of the Nuclear Envelope and Associated

Wei Xie, Henning F. Horn, and Graham D. Wright

  1. Analyses of the Dynamic Properties of Nuclear Lamins by Fluorescence Recovery after
    Photobleaching (FRAP) and Fluorescence Correlation Spectroscopy (FCS)

    Shimi Takeshi, Chan-Gi
    Pack, and Robert D. Goldman

  2. Probing Protein
    Distribution Along the Nuclear Envelope In Vivo by Using Single-Point FRAP

    Krishna C. Mudumbi and Weidong Yang

  3. The Use of
    2-Photon FLIM/FRET to Study Protein Interactions during Nuclear Envelope
    Fusion In Vivo and In Vitro

    Richard D. Byrne, Banafsh
    é Larijani, and Dominic L. Poccia

  4. Identifying
    Protein-Protein Associations at the Nuclear Envelope with BioID

    In Kim, Samuel C. Jensen, and Kyle J. Roux

  5. In Situ
    Detection of Interactions between Nuclear Envelope Proteins and Partners

    Alice Barateau and Brigitte Buendia

  6. Methods for
    Single-Cell Pulse-Chase Analysis of Nuclear Components

    Drozdz and David J. Vaux

  7. Analysis of
    Nuclear Lamina Proteins in Myoblast Differentiation by Functional Complementation

    Tapia and Larry Gerace

  8. Analysis of
    Meiotic Telomere Behavior in the Mouse

    Link, Ricardo Benavente, and Manfred Alsheimer

III Nuclear Envelope Interactions with the Cytoskeleton

  1. Identification and Validation of Putative Nesprin Variants

    Flavia Autore,
    Catherine M. Shanahan, and Qiuping Zhang

  2. Detection of Diverse and High Molecular Weight Nesprin-1 and Nesprin-2 Isoforms Using
    Western Blotting

    James Carthew and Iakowos

  3. The
    Use of Polyacrylamide Hydrogels to Study the Effects of Matrix Stiffness on Nuclear Envelope Properties

    Minaisah, Susan Cox, and Derek T. Warren

  4. Cell Microharpooning to Study Nucleo-Cytoskeletal Coupling

    Fedorchak and Jan Lammerding

  5. Wound
    Healing Assays to Study Mechanisms of Nuclear Movement in Fibroblasts and Myoblasts

    Wakam Chang, Susumu Antoku, and
    Gregg G. Gundersen

  6. Methods for
    Assessing Nuclear Rotation and Nuclear Positioning in Developing Skeletal Muscle

    Meredith H. Wilson, Matthew G. Bray, and Erika L. F. Holzbaur

  7. Imaging Approaches to Investigate Myonuclear Positioning in Drosophila

    Mafalda Azavedo, Victoria K. Schulman,
    Eric Folker, Mridula Balakrishnana, and Mary Baylies

IV Nuclear Envelope-Chromatin Interactions

    Nuclear Lamin-Genome Interactions by Chromatin
    Immunoprecipitation of Nuclear Lamins

    Anja R. Oldenburg and
    Philippe Collas

  1. Lamin ChIP
    from Chromatin Prepared by Micrococcal Nuclease Digestion

    Isabelle Dubant-Goulet

  2. DamID
    Analysis of Nuclear Organization in Caenorhabditis elegans

    Georgina G
    Peter Meister, and Peter Askjaer

  3. The
    Application of DamID to Identify Peripheral Gene Sequences in
    Differentiated and Primary Cells

    Michael I. Robson and
    Eric C. Schirmer

  4. Visualizing the Spatial Relationship of the Genome with the
    Nuclear Envelope Using Fluorescence in situ Hybridization

    Craig S. Clements, Ural Bikkul,
    Mai Hassan Ahmed, Helen A. Foster, Lauren S. Godwin, and Joanna M. Bridger

  5. Visualization of Genomic Loci in Living Cells with a Fluorescent CRISPR/Cas9 System

    Tobias Anton, Heinrich
    Leonhardt, and Yolanda Markaki

  6. Methods to
    Monitor DNA Repair Defects and Genomic Instability in the Context of a
    Disrupted Nuclear Lamina

    Suzanna Gonzalo and Ray Kreienkamp

V Nucleo-Cytoplasmic Transport

  1. High Resolution
    Scanning Electron Microscopy and Immuno-Gold Labeling of the Nuclear Lamina and Nuclear Pore Complex

    Martin W. Goldberg

  2. An In Vitro Assay to Study Targeting of Membrane Proteins to the Inner Nuclear Membrane

    Ungricht, Sumit Pawar, and Ulrike Kutay

  3. Nuclear Protein
    Transport in Digitonin Permeabilized Cells

    Steven A. Adam

  4. Analysis of
    CRM1-Dependent Nuclear Export in Permeabilized Cells

    Ralph H. Kehlenbach and
    Sarah A. Port

  5. SPEED Microscopy and Its Application in Nucleocytoplasmic Transport

    Jiong Ma, Joseph M. Kelich, and Weidong Yang

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