Immunocytochemical Methods and Protocols

1. Front Matter

   Pages i-xii

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2. Overview of Antibody Use in Immunocytochemistry

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

   Pages 3-11

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

   • Ute M. Kent

   Pages 13-21

4. Purification of Antibodies Using Ion-Exchange Chromatography

   • Ute M. Kent

   Pages 23-27

5. Purification of Antibodies Using Affinity Chromatography

   • Ute M. Kent

   Pages 29-35

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

   • Ute M. Kent

   Pages 37-41

7. Conjugation of Fluorochromes to Antibodies

   • Su-Yau Mao

   Pages 43-47

8. Biotinylation of Antibodies

   • Su-Yau Mao

   Pages 49-52

9. Overview of Cell Fixation and Permeabilization

   • Melissa A. Melan

   Pages 55-66

10. Preparation of Frozen Sections for Analysis

    • Gary L. Bratthauer

    Pages 67-73

11. Processing of Cell-Touch Preparations

    • Gary L. Bratthauer

    Pages 75-79

12. Processing of Cytological Specimens

    • Gary L. Bratthauer

    Pages 81-87

13. Processing of Tissue-Culture Cells

    • Gary L. Bratthauer

    Pages 89-95

14. Processing of Tissue Specimens

    • Gary L. Bratthauer

    Pages 97-104

15. Overview of Fluorophores

    • J. Michael Mullins

    Pages 107-116

16. Direct Immunofluorescent Labeling of Cells

    • Lorette C. Javois

    Pages 117-121

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

    • Mark C. Willingham

    Pages 123-130

18. Fluorescence Labeling of Intracellular Antigens of Attached or Suspended Tissue-Culture Cells

    • Mark C. Willingham

    Pages 131-142

19. Fluorescent Labeling of Surface or Intracellular Antigens in Whole-Mounts

    • Lorette Javois

    Pages 143-153

20. Overview of Antigen Detection Through Enzymatic Activity

    • Gary L. Bratthauer

    Pages 155-164

The principle that antibodies can be used as cytochemical agents provided they are tagged with suitable markers has been evident for over 50 years. During this time the use of immunocytochemical meth­ ods has spread to a wide array of biological disciplines. Early applica­ tions focused on the detection of microbial antigens in tissues, while more recent applications have used monoclonal antibodies to study cell differentiation during embryonic development. For a select few disci­ plines, volumes have been published focusing on the specific applica­ tion of immunocytochemical techniques to that discipline. What distinguishes the present book, Immunocytochemical Meth­ ods and Protocols, from earlier books is its broad appeal to researchers in all disciplines, including those in both research and clinical settings. The methods and protocols presented here are designed to be general in their application and the accompanying "Notes" provide invaluable assistance in adapting or troubleshooting the protocols. Interspersed throughout the book are chapters providing overviews of select topics related to immunocytochemistry.

This book is a useful, sometimes beautiful, compilation of protocols and methods...-FEBS Letters
...offer[s[ unique features that make [it a] worthwhile purchase...comprehensive-Trends in Genetics

• The Catholic University of America, Washington, DC

  Lorette C. Javois

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