Beschreibung

This third edition reflects the rapid evolution of immunocytochemistry in the post-genomic era. Expert researchers explore the latest methods, now widely used in situ to identify components of cells and tissues in both normal and pathological conditions.

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.

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

Klappentext

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.

Inhalt
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

Produktinformationen

Titel:	Immunocytochemical Methods and Protocols
Editor:	Maria Célia Jamur   Constance Oliver
EAN:	9781588294630
ISBN:	1588294633
Format:	Fester Einband
Herausgeber:	Humana Press
Genre:	Medizin
Anzahl Seiten:	432
Gewicht:	1005g
Größe:	H260mm x B183mm x T28mm
Jahr:	2009
Untertitel:	Englisch
Auflage:	3rd ed. 2010