Immunocytochemical Methods and Protocols

1. Front Matter

   Pages i-xii

   PDF

2. Antibody Preparation

   1. Front Matter

      Pages 1-1

      PDF

   2. Overview of Antibody Use in Immunocytochemistry

      • Su-Yau Mao, Lorette C. Javois, Ute M. Kent

      Pages 3-10

   3. Purification of Antibodies Using Ammonium Sulfate Fractionation or Gel Filtration

      • Ute M. Kent

      Pages 11-18

   4. Purification of Antibodies Using Ion-Exchange Chromatography

      • Ute M. Kent

      Pages 19-22

   5. Purification of Antibodies Using Affinity Chromatography

      • Ute M. Kent

      Pages 23-28

   6. Purification of Antibodies Using Protein A-Sepharose and FPLC

      • Ute M. Kent

      Pages 29-33

   7. Conjugation of Fluorochromes to Antibodies

      • Su-Yau Mao

      Pages 35-38

   8. Biotinylation of Antibodies

      • Su-Yau Mao

      Pages 39-41

3. Tissue Preparation for Light Microscopic Analyses

   1. Front Matter

      Pages 43-43

      PDF

   2. Overview of Cell Fixatives and Cell Membrane Permeants

      • Melissa A. Melan

      Pages 45-55

   3. Preparation of Frozen Sections for Analysis

      • Gary L. Bratthauer

      Pages 57-62

   4. Processing of Cytological Specimens

      • Gary L. Bratthauer

      Pages 63-70

   5. Processing of Tissue-Culture Cells

      • Gary L. Bratthauer

      Pages 71-76

   6. Processing of Tissue Specimens

      • Gary L. Bratthauer

      Pages 77-84

   7. Antigen Retrieval for Immunohistochemical Reactions in Routinely Processed Paraffin Sections

      • Laszlo Krenacs, Tibor Krenacs, Mark Raffeld

      Pages 85-93

4. Light Microscopic Detection Systems

   1. Front Matter

      Pages 95-95

      PDF

   2. Overview of Fluorochromes

      • J. Michael Mullins

      Pages 97-105

   3. Direct Immunofluorescent Labeling of Cells

      • Lorette C. Javois

      Pages 107-111

   4. Fluorescence Labeling of Surface Antigens of Attached or Suspended Tissue-Culture Cells

      • Mark C. Willingham

      Pages 113-119

Lorette Javois' timely new 2nd edition revises and updates her widely acclaimed collection of step-by-step immunocytochemical methods, one that is now used in many biological and biomedical research programs. The methods are designed for researchers and clinicians who wish to visualize molecules in plant or animal embryos, tissue sections, cells, or organelles. In addition to cutting-edge protocols for purifying and preparing antibodies, light microscopic analysis, confocal microscopy, FACS, and electron microscopy, this revised edition contains many new methods for applying immunocytochemical techniques in the clinical laboratory and in combination with in situ hybridization.

• DNA
• ELISA
• Filtration
• Organelle
• PCR
• cell membrane
• chemistry
• electron microscopy
• hybridization
• membrane
• metabolism
• microscopy
• nucleic acid
• protein
• tissue

"This second edition provides detailed protocols for immunocytochemical localization of tissue constituents. The book is well balanced and suited for basic and clinical research scientists. . .The editor has successfully compiled this book for the intended readers. The editor and contributors are credible. . .The protocol style format is useful for a first experiment. The illustrations are attractive and informative. The book as a whole is nicely done. . .I find this a very useful and practical addition to all fields of biology and medicine. It is very user friendly and should be a great reference for those designing experiments and writing."-Doody's Health Sciences Book Review Journal

"...this excellent book on Methods and Protocols can be recommended to both investigators starting out in the field of immunocytochemistry and to basic and clinical researchers already working in the field because it contains a plethora of methods and protocols for the demonstration of moleculesin many different normal and pathological situations and also under different experimental conditions." - Journal of Chemical Neuroanatomy

"...this book would make a valuable addition to laboratories currently involved in research using immunocytochemical techniques and would be particularly useful for those wishing to introduce these techniques into their research programmes...Given the depth of coverage of theory behind various techniques and the detailed notes that accompany each set of protocols, the book would also make an ideal teaching aid."-Molecular Biotechnology

• Catholic University of America, Washington

  Lorette C. Javois

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