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Immunocytochemical Methods and Protocols

Series: Methods in Molecular Biology

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by Constance Oliver and Maria Celia Jamur

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Immunocytochemical Methods and Protocols

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Main description:

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.

Feature:

Fully updated from the first edition to offer cutting-edge methods in immunocytochemistry

Includes methods applicable for the basic research laboratory as well as the clinical laboratory

Provides the theoretical basis behind the methods

Contains methods that have multidisciplinary applications

Suitable for beginning and experienced investigators

Back cover:

The landscape of immunocytochemistry has significantly evolved in past years. Once a laborious technique with limited application, the introduction of new detection systems and instrumentation has facilitated the widespread use of immunocytochemical methods. In the post-genomic era, the use of immunocytochemical methods has grown to include the identification of a broad spectrum of antigens including proteins, carbohydrates and lipids from virtually every organism. In Immunocytochemical Methods and Protocols, Third Edition, expert researchers explore the modern methods employed in the field today, now widely used to identify, in situ, various components of cells and tissues in both normal and pathological conditions, and to demonstrate the localization of subcellular components. Chapters provide methods that are designed to be general in their application and to appeal to investigators across all disciplines, including those in research and clinical settings. Composed in the highly successful Methods in Molecular Biology™ series format, chapters contain a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls.

Comprehensive and ground-breaking, Immunocytochemical Methods and Protocols, Third Edition, brings together a variety of methods that are normally limited to applications in a specific area, providing a unique and vital guide for researchers and clinicians alike.

Contents:

Immunocytochemical Methods and Protocols, Third Edition
Constance Oliver and Maria Célia Jamur, Eds.

Table of Contents

Preface

Contributors

Part I. Antibody Preparation

Overview of Antibodies for Immunochemistry

Constance Oliver and Maria Célia Jamur

Introduction to the Purification of Antibodies

Ana Cristina Grodzki and Elsa Berenstein

Antibody Purification: Ammonium Sulfate Fractionation or Gel Filtration

Ana Cristina Grodzki and Elsa Berenstein

Antibody Purification: Ion - Exchange chromatography

Ana Cristina Grodzki and Elsa Berenstein

Antibody Purification: Affinity Chromatography - Protein A and Protein G Sepharose

Ana Cristina Grodzki and Elsa Berenstein

Conjugation of Fluorochromes to Antibodies

Su-Yau Mao

Biotinylation of Antibodies

Su-Yau Mao

Part II. Tissue Preparation for Light Microscopic Analysis

Cell Fixatives for Immunostaining

Maria Célia Jamur and Constance Oliver

Permeablization of Cell Membranes

Maria Célia Jamur and Constance Oliver

Preparation of Frozen Sections for Analysis

Gary L. Bratthauer

Processing of Cytological Specimens

Gary L. Bratthauer

Processing of Tissue Culture Cells

Gary L. Bratthauer

Processing Of Tissue Specimens

Gary L. Bratthauer

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

Laszlo Krenacs, Tibor Krenacs, Eva Stekovics,and Mark Raffeld

Part III: Bright Field Detection Systems

Fluorochromes - Properties and Characteristics

J. Michael Mullins

Direct Immunofluorescent Labeling of Cells

Maria Veronica Dávila Pástor

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

Mark C. Willingham

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

Mark C. Willingham

Fluorescent Visualization of Macromolecules in Drosophila Whole Mounts

Ricardo Guelerman Pinheiro Ramos, Luciana Claudia Herculano Machado, and Livia Maria Rosatto Moda

Overview of Conventional Fluorescence Photomicrography

J. Michael Mullins

Overview of Confocal Microscopy

William D. Swaim

Overview of Laser Microbeam Applications as Related to Antibody Targeting

P. Scott Pine

Immuno-Laser Capture Microdissection of Rat Brain Neurons For Real Time Quantitative PCR

L. Scot Bastian and Denis G. Baskin

Overview of Antigen Detection through Enzymatic Activity

Gary L. Bratthauer

The Peroxidase-antiperoxidase (PAP) Method and Other All-Immunologic Detection Methods

Gary L. Bratthauer

The Avidin-Biotin Complex (ABC) Method and Other Avidin-Biotin Binding Methods

Gary L. Bratthauer

Avidin-Biotin Labeling of Cellular Antigens in Cryostat-Sectioned Tissue

Mark Raffeld and Elaine S. Jaffe

Multiple Antigen Immunostaining Procedures

Tibor Krenacs, Laszlo Krenacs and Mark Raffeld

Immunoenzymatic Quantitative Analysis of Antigens

PRODUCT DETAILS

ISBN-13: 9781588294630
Publisher: Springer (Humana Press)
Publication date: December, 2009
Pages: 416
Weight: 1000g
Availability: Not available (reason unspecified)
Subcategories: Immunology, Pathology

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CUSTOMER REVIEWS

Average Rating

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)