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High-Resolution Imaging of Cellular Proteins

Methods and Protocols

  • Book
  • © 2016

Overview

Editors:
  1. Steven D. Schwartzbach

    1. Department of Biological Sciences, The University of Memphis, Memphis, USA

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    You can also search for this editor in PubMed   Google Scholar

  2. Omar Skalli

    1. Department of Biological Sciences, The University of Memphis, Memphis, USA

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

  3. Thomas Schikorski

    1. Department of Anatomy, Universidad Central del Caribe, Bayamon, USA

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

  • Includes cutting-edge methods and protocols
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice from the experts
  • Includes supplementary material: sn.pub/extras

Part of the book series: Methods in Molecular Biology (MIMB, volume 1474)

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Table of contents (22 protocols)

  1. Front Matter

    Pages i-xiii
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  2. Molecular Toolbox

    1. Front Matter

      Pages 1-1
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    2. Expression of Epitope-Tagged Proteins in Mammalian Cells in Culture

      • Jay M. Bhatt, Melanie L. Styers, Elizabeth Sztul
      Pages 3-24

    3. Antibody Production with Synthetic Peptides

      • Bao-Shiang Lee, Jin-Sheng Huang, Lasanthi P. Jayathilaka, Jenny Lee, Shalini Gupta
      Pages 25-47

    4. Production and Purification of Polyclonal Antibodies

      • Masami Nakazawa, Mari Mukumoto, Kazutaka Miyatake
      Pages 49-59

    5. Preparation of Colloidal Gold Particles and Conjugation to Protein A/G/L, IgG, F(ab′)2, and Streptavidin

      • Sadaki Yokota
      Pages 61-71

    6. Helper-Dependent Adenoviral Vectors and Their Use for Neuroscience Applications

      • Mónica S. Montesinos, Rachel Satterfield, Samuel M. Young Jr.
      Pages 73-90

  3. Fluorescent Microscopy Toolbox

    1. Front Matter

      Pages 91-91
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    2. Localizing Proteins in Fixed Giardia lamblia and Live Cultured Mammalian Cells by Confocal Fluorescence Microscopy

      • Lilian Nyindodo-Ogari, Steven D. Schwartzbach, Omar Skalli, Carlos E. Estraño
      Pages 93-111

    3. Using Fluorescent Protein Fusions to Study Protein Subcellular Localization and Dynamics in Plant Cells

      • Yong Cui, Caiji Gao, Qiong Zhao, Liwen Jiang
      Pages 113-123

    4. Using FRAP or FRAPA to Visualize the Movement of Fluorescently Labeled Proteins or Cellular Organelles in Live Cultured Neurons Transformed with Adeno-Associated Viruses

      • Yaara Tevet, Daniel Gitler
      Pages 125-151

    5. Bimolecular Fluorescence Complementation (BiFC) Analysis of Protein–Protein Interactions and Assessment of Subcellular Localization in Live Cells

      • Evan P. S. Pratt, Jake L. Owens, Gregory H. Hockerman, Chang-Deng Hu
      Pages 153-170

    6. Viral Injection and Cranial Window Implantation for In Vivo Two-Photon Imaging

      • Gordon B. Smith, David Fitzpatrick
      Pages 171-185

    7. Imaging Synaptic Vesicle Exocytosis-Endocytosis with pH-Sensitive Fluorescent Proteins

      • Olusoji A. T. Afuwape, Ege T. Kavalali
      Pages 187-200

  4. Electron Microscopy Toolbox

    1. Front Matter

      Pages 201-201
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    2. Immunogold Protein Localization on Grid-Glued Freeze-Fracture Replicas

      • Harumi Harada, Ryuichi Shigemoto
      Pages 203-216

    3. Pre-embedding Double-Label Immunoelectron Microscopy of Chemically Fixed Tissue Culture Cells

      • Lou G. Boykins, Jonathan C. R. Jones, Carlos E. Estraño, Steven D. Schwartzbach, Omar Skalli
      Pages 217-232

    4. Immunoelectron Microscopy of Cryofixed and Freeze-Substituted Plant Tissues

      • Miyuki Takeuchi, Keiji Takabe, Yoshinobu Mineyuki
      Pages 233-242

    5. Immunoelectron Microscopy of Cryofixed Freeze-Substituted Yeast Saccharomyces cerevisiae

      • Jindřiška Fišerová, Christine Richardson, Martin W. Goldberg
      Pages 243-258

    6. Pre-embedding Method of Electron Microscopy for Glycan Localization in Mammalian Tissues and Cells Using Lectin Probes

      • Yoshihiro Akimoto, Kuniaki Takata, Hayato Kawakami
      Pages 259-267
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Keywords

About this book

This volume presents authoritative and cutting-edge methods and protocols focusing on three tool boxes covering the increasingly diverse methodologies used to image selected proteins and to investigate their function by light and electron microscopy. The first tool box includes the development of a wide range of molecular and immunological probes to target specific proteins. The second details the use of these probes for high resolution fluorescence microscopy and the third focuses on applications for transmission and scanning electron microscopy. 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 thus ensuring successful results in the further study of this vital field.

Reviews

“This is a collection of timely techniques and methods that have been key for the development of imaging at the structural and ultrastructural level. … The intended audience is cell biologists and all scientists working with imaging techniques to localize protein expression at a cellular and subcellular level. … Each chapter ends with a list of references. Graphics and high-quality figures are displayed where needed.” (Michele Fornaro, Doody's Book Reviews, July, 2017)

Editors and Affiliations

  • Department of Biological Sciences, The University of Memphis, Memphis, USA

    Steven D. Schwartzbach, Omar Skalli

  • Department of Anatomy, Universidad Central del Caribe, Bayamon, USA

    Thomas Schikorski

Bibliographic Information

  • Book Title: High-Resolution Imaging of Cellular Proteins

  • Book Subtitle: Methods and Protocols

  • Editors: Steven D. Schwartzbach, Omar Skalli, Thomas Schikorski

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-4939-6352-2

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media New York 2016

  • Hardcover ISBN: 978-1-4939-6350-8Published: 12 August 2016

  • Softcover ISBN: 978-1-4939-8173-1Published: 09 June 2018

  • eBook ISBN: 978-1-4939-6352-2Published: 11 August 2016

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XIII, 366

  • Number of Illustrations: 24 b/w illustrations, 21 illustrations in colour

  • Topics: Imaging / Radiology

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