Antigen Retrieval Immunohistochemistry Based Research and Diagnostics / Edition 1

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The most complete, up-to-date reference on antigen retrieval and immunohistochemistry An antigen is a substance that prompts the generation of antibodies and can cause an immune response. The antigen retrieval (AR) technique is in wide use across the globe, and is a critical technique used in medical diagnosis of disease, particularly clinical targeted cancer treatment. Antigen Retrieval Immunohistochemistry Based Research and Diagnostics discusses several scientific approaches to the standardization of quantifiable immunohistochemistry (IHC). Based on the development and application of AR by the editors, this volume summarizes recent achievements in AR-IHC and analyzes numerous cutting-edge issues for future research projects. Featuring contributions from a worldwide group of leading experts and research scientists in the field, this important work:
• Summarizes the key problems in the four fields of antigen retrieval
• Discusses the advances of AR techniques and their applications
• Provides practical methods and protocols in AR-IHC, such as extraction of nucleic acids and proteins for molecular analysis, cell/tissue sample preparation, and standardization and development of various techniques to meet the future needs of clinical and research molecular analysis
• Encourages further research in AR and IHC, particularly how AR methods might be employed for improved test performance and the development of greater reliability and reproducibility of IHC
• Includes an appendix of related laboratory protocols Antigen Retrieval Immunohistochemistry Based Research and Diagnostics is intended for clinical pathologists, molecular cell biologists, basic research scientists, technicians, and graduate students who undertake tissue/cell morphologic and molecular analysis and wish to use and extend the power of immunohistochemistry. It is also pertinent for most biotechnology companies majoring in development of IHC products.

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Product Details

Meet the Author

Shan-Rong Shi, MD, is professor of clinical pathology at theKeck School of Medicine of the University of Southern California.He is the Associate Editor of The Journal of Histochemistry andcytochemistry, he is on the Editoral Board of AppliedImmunohistochemistry and Molecular Morphology and is also on theOverseas Editorial Board of the Chinese Journal of Pathology. Dr.Shi is the ineventor of the antigen retrieval (AR) technique.

Clive R. Taylor, MD, PhD, is Senior Associate Dean,Professor and Chairman of Pathology at the Keck School of Medicine.Dr. Taylor is Editor in Chief of Applied Immunohistochemistry andMolecular Morphology. Dr. Taylor is a world renowned expert inimmunohistochemistry and clinical pathology.

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Table of Contents


1 Standardization of Antigen Retrieval Techniques Based on the Test Battery Approach Clive R. Taylor Taylor, Clive R. 3

2 Extended Application of Antigen Retrieval Technique in Immunohistochemistry and in Situ Hybridization Clive R. Taylor Taylor, Clive R. 25

3 Extraction of DNA/RNA From Formalin-Fixed, Paraffin-Embedded Tissue Based on the Antigen Retrieval Principle Clive R. Taylor Taylor, Clive R. 47


4 Key Issues and Strategies of Standardization for Quantifiable Immunohistochemistry Clive R. Taylor Taylor, Clive R. 75

5 Standardization of Immunohistochemistry Based on Antigen Retrieval Technique Clive R. Taylor Taylor, Clive R. 87

6 Standard Reference Material: Cell Line Development and Use of Reference Cell Lines as Standards For External Quality Assurance of HER2 IHC and ISH Testing Keith Miller Miller, Keith 101

7 Peptides as Immunohistochemistry Controls Seshi R. Sompuram Sompuram, Seshi R. 123

8 Standard Reference Material: Proteinembedding Technique and Design of Bar Code Clive R. Taylor Taylor, Clive R. 141

9 The Pros and Cons of Automation For Immunohistochemistry From the Prospective of the Pathology Laboratory Loralee McMahon McMahon, Loralee 151

10 Image Analysis in Immunohistochemistry Alton D. Floyd Floyd, Alton D. 165


11 Tissue Cell Sample Preparation: Lessons from the Antigen Retrieval Technique Clive R. Taylor Taylor, Clive R. 189

12 Mechanisms of Action and Proper Use of Common fixatives Richard W. Dapson Dapson, Richard W. 195

13 Cell Sample Preparation For Clinical Cytopathology: Current Status and Future Development Patricia G. Wasserman Wasserman, Patricia G. 219

14 Design of a Tissue Surrogate to Examine Accuracy of Proteomic Analysis Timothy J. O'Leary O'Leary, Timothy J. 235


15 Study of Formalin Fixation and Heat-Induced Antigen Retrieval Timothy J. O'Leary O'Leary, Timothy J. 253

16 A Linear Epitopes Model of Antigen Retrieval Seshi R. Sompuram Sompuram, Seshi R. 287

17 pH or Ionic Strength of Antigen Retrieval Solution: A Potential Role for Refolding During Heat Treatment Shuji Yamashita Yamashita, Shuji 303

18 Commentary: Future Directions Jeffrey T. Mason Mason, Jeffrey T. 323


19 Techniques of Protein Extraction from FFPE Tissue/Cells for Mass Spectrometry Jeffrey T. Mason Mason, Jeffrey T. 335

20 Application of Shotgun Proteomics to Formalin-Fixed and Paraffin-Embedded Tissues Brian M. Balgley Balgley, Brian M. 347

21 Visualizing Protein Maps In Tissue Mitsutoshi Setou Setou, Mitsutoshi 369

22 Symbiosis of Immunohistochemistry and Proteomics: Marching to a New Era Clive R. Taylor Taylor, Clive R. 391

Appendix Related Laboratory Protocols 399

Index 413

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