Immunocytochemical Methods and Protocols

1. Front Matter

   Pages i-xii

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2. Antibody Preparation

   1. Front Matter

      Pages 1-1

      PDF

   2. Overview of Antibodies for Immunochemistry

      • Constance Oliver, Maria Célia Jamur

      Pages 3-9

   3. Introduction to the Purification of Antibodies

      • Ana Cristina Grodzki, Elsa Berenstein

      Pages 11-13

   4. Antibody Purification: Ammonium Sulfate Fractionation or Gel Filtration

      • Ana Cristina Grodzki, Elsa Berenstein

      Pages 15-26

   5. Antibody Purification: Ion-Exchange Chromatography

      • Ana Cristina Grodzki, Elsa Berenstein

      Pages 27-32

   6. Antibody Purification: Affinity Chromatography – Protein A and Protein G Sepharose

      • Ana Cristina Grodzki, Elsa Berenstein

      Pages 33-41

   7. Conjugation of Fluorochromes to Antibodies

      • Su-Yau Mao, J. Michael Mullins

      Pages 43-48

   8. Biotinylation of Antibodies

      • Su-Yau Mao

      Pages 49-52

3. Tissue Preparation for Light Microscopic Analysis

   1. Front Matter

      Pages 54-54

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   2. Cell Fixatives for Immunostaining

      • Maria Célia Jamur, Constance Oliver

      Pages 55-61

   3. Permeabilization of Cell Membranes

      • Maria Célia Jamur, Constance Oliver

      Pages 63-66

   4. Preparation of Frozen Sections for Analysis

      • Gary L. Bratthauer

      Pages 67-73

   5. Processing of Cytological Specimens

      • Gary L. Bratthauer

      Pages 75-84

   6. Processing of Tissue Culture Cells

      • Gary L. Bratthauer

      Pages 85-92

   7. Processing of Tissue Specimens

      • Gary L. Bratthauer

      Pages 93-102

   8. Heat-Induced Antigen Retrieval for Immunohistochemical Reactions in Routinely Processed Paraffin Sections

      • Laszlo Krenacs, Tibor Krenacs, Eva Stelkovics, Mark Raffeld

      Pages 103-119

4. Bright Field Detection Systems

   1. Front Matter

      Pages 122-122

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   2. Fluorochromes: Properties and Characteristics

      • J. Michael Mullins

      Pages 123-134

   3. Direct Immunofluorescent Labeling of Cells

      • Maria Veronica Dávila Pástor

      Pages 135-142

Antibodies tagged with fuorescent markers have been used in histochemistry for over 50 years. Although early applications were focused on the detection of microbial antigens in tissues, the use of immunocytochemical methods now has spread to include the det- tion of a wide array of antigens including proteins, carbohydrates, and lipids from virtually any organism. Today, immunohistochemistry is widely used to identify, in situ, various components of cells and tissues in both normal and pathological conditions. The method gains its strength from the extremely sensitive interaction of a specifc antibody with its antigen. For some scientifc areas, books have been published on applications of immu- cytochemical techniques specifc to that area. What distinguished Immunocytochemical Methods and Protocols from earlier books when it was frst published was its broad appeal to investigators across all disciplines, including those in both research and clinical settings. The methods and protocols p- sented in the frst edition were designed to be general in their application; the accompa- ing “Notes” provided the reader with invaluable assistance in adapting or troubleshooting the protocols. These strengths continued to hold true for the second edition and again for the third edition. Since the publication of the frst edition, the application of immuno- tochemical techniques in the clinical laboratory has continued to rise and this third edition provides methods that are applicable to basic research as well as to the clinical laboratory.

• Antibody
• Antigen
• Fluorescence microscopy
• IgG
• Immunohistochemistry
• Immunostaining

From the reviews of the third edition:

“This book targets not only immunologists but a very broad readership. … very well-written and will be equally useful for a complete beginner to the technique, as well as for a scientist who has been using the methods for years. … I would recommend this book in particular to researchers who start to use immunocyto/histochemistry in their work … . The book should also be considered by laboratories that would like to have a reference for reliable immunostaining protocols.” (Anna Vossenkaemper, Immunology News, August, 2010)

• Fac. Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil

  Constance Oliver, Maria Célia Jamur

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