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Immunoelectron Microscopy

Methods and Protocols

  • Book
  • © 2010

Overview

Editors:
  1. Steven D. Schwartzbach

    1. Dept. Biology, University of Memphis, Memphis, USA

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    You can also search for this editor in PubMed   Google Scholar

  2. Tetsuaki Osafune

    1. Dept. Life Science, Nippon Sport Science University, Yokohama, Japan

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

  • Presents a wide range of Cryo and chemical fixation methods for single cells, plant, and animal tissue
  • Contains Immunogold labeling methods for transmission and scanning electron microscopy
  • Includes comprehensive pre and post embedding immunolabeling methods
  • Includes supplementary material: sn.pub/extras

Part of the book series: Methods in Molecular Biology (MIMB, volume 657)

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Table of contents (26 protocols)

  1. Front Matter

    Pages i-xi
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  2. Molecular Toolbox

    1. Front Matter

      Pages 1-1
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    2. Protein Antigen Expression in Escherichia coli for Antibody Production

      • David M. Rancour, Steven K. Backues, Sebastian Y. Bednarek
      Pages 3-20

    3. Expression of Epitope-Tagged Proteins in Plants

      • Takuya Furuichi
      Pages 21-31

    4. Expression of Epitope-Tagged Proteins in Arabidopsis Leaf Mesophyll Protoplasts

      • Young-Hee Cho, Sang-Dong Yoo
      Pages 33-42

    5. Transient Expression of Epitope-Tagged Proteins in Mammalian Cells

      • Melanie L. Styers, Jason Lowery, Elizabeth Sztul
      Pages 43-61

    6. Production and Purification of Polyclonal Antibodies

      • Masami Nakazawa, Mari Mukumoto, Kazutaka Miyatake
      Pages 63-74

    7. Production and Purification of Monoclonal Antibodies

      • Masami Nakazawa, Mari Mukumoto, Kazutaka Miyatake
      Pages 75-91

    8. Production of Antipeptide Antibodies

      • Bao-Shiang Lee, Jin-Sheng Huang, G. D. Lasanthi P. Jayathilaka, Syed S. Lateef, Shalini Gupta
      Pages 93-108

    9. Preparation of Colloidal Gold Particles and Conjugation to Protein A, IgG, F(ab’)2, and Streptavidin

      • Sadaki Yokota
      Pages 109-119

  3. Microscopy Toolbox

    1. Front Matter

      Pages 121-121
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    2. Immunoelectron Microscopy of Chemically Fixed Developing Plant Embryos

      • Tetsuaki Osafune, Steven D. Schwartzbach
      Pages 123-132

    3. Pre-embedding Immunogold Localization of Antigens in Mammalian Brain Slices

      • Thomas Schikorski
      Pages 133-144

    4. Pre-embedding Immunoelectron Microscopy of Chemically Fixed Mammalian Tissue Culture Cells

      • Haruo Hagiwara, Takeo Aoki, Takeshi Suzuki, Kuniaki Takata
      Pages 145-154

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

      • Miyuki Takeuchi, Keiji Takabe, Yoshinobu Mineyuki
      Pages 155-165

    6. In Vivo Cryotechniques for Preparation of Animal Tissues for Immunoelectron Microscopy

      • Shinichi Ohno, Nobuhiko Ohno, Nobuo Terada, Sei Saitoh, Yurika Saitoh, Yasuhisa Fujii
      Pages 167-179

    7. Immunoelectron Microscopy of Cryofixed Freeze-Substituted Mammalian Tissue Culture Cells

      • Akira Sawaguchi
      Pages 181-190

    8. Immunoelectron Microscopy of Cryofixed Freeze-Substituted Saccharomyces cerevisiae

      • Jindriska Fiserova, Martin W. Goldberg
      Pages 191-204

    9. High-Resolution Molecular Localization by Freeze-Fracture Replica Labeling

      • Akikazu Fujita, Toyoshi Fujimoto
      Pages 205-216

    10. Pre-embedding Electron Microscopy Methods for Glycan Localization in Chemically Fixed Mammalian Tissue Using Horseradish Peroxidase-Conjugated Lectin

      • Yoshihiro Akimoto, Hayato Kawakami
      Pages 217-224
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Keywords

About this book

Immunoelectron microscopy is a key technique that bridges the information gap between biochemistry, molecular biology, and ultrastructural studies placing macromolecular functions within a cellular context. In Immunoelectron Microscopy: Methods and Protocols, expert researchers combine the tools of the molecular biologist with those of the microscopist. From the molecular biology toolbox, this volume presents methods for antigen production by protein expression in bacterial cells, methods for epitope tagged protein expression in plant and animal cells allowing protein localization in the absence of protein specific antibodies as well as methods for the production of anti-peptide, monoclonal, and polyclonal antibodies. From the microscopy toolbox, sample preparation methods for cells, plant, and animal tissue are presented. Both cryo-methods, which have the advantage of retaining protein antigenicity at the expense of ultrastructural integrity, as well as chemical fixation methods that maintain structural integrity while sacrificing protein antigenicity have been included, with chapters examining various aspects of immunogold labeling. 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 notes on troubleshooting and avoiding known pitfalls. Authoritative and essential, Immunoelectron Microscopy: Methods and Protocols seeks to facilitate an increased understanding of structure function relationships.

Reviews

From the reviews:

“The editors tell us that Immunoelectron microscopy is the technique that bridges the information gap between biochemistry, molecular biology, and ultrastructural studies placing macromolecular functions within a cellular context. Immunoelectron microscopy can be used on virtually every unicellular and multicellular organism. … An excellent source of information about the minutiae of preparation techniques if this is your area.” (P. W. Hawles, Ultramicroscopy, Vol. 111 (7), June, 2011)

Editors and Affiliations

  • Dept. Biology, University of Memphis, Memphis, USA

    Steven D. Schwartzbach

  • Dept. Life Science, Nippon Sport Science University, Yokohama, Japan

    Tetsuaki Osafune

Bibliographic Information

  • Book Title: Immunoelectron Microscopy

  • Book Subtitle: Methods and Protocols

  • Editors: Steven D. Schwartzbach, Tetsuaki Osafune

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-60761-783-9

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC 2010

  • Hardcover ISBN: 978-1-60761-782-2Published: 20 July 2010

  • Softcover ISBN: 978-1-4939-5758-3Published: 23 August 2016

  • eBook ISBN: 978-1-60761-783-9Published: 05 July 2010

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XI, 352

  • Topics: Cell Biology, Immunology, Antibodies

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