Immunocytochemical Methods and Protocols [electronic resource] / edited by Constance Oliver, Maria Célia Jamur.

Totowa, NJ : Humana Press, 2010.
Methods in Molecular Biology, Methods and Protocols, 1064-3745 ; 588
Springer Protocols (Springer-12345)
Methods in Molecular Biology, Methods and Protocols, 1064-3745 ; 588
1 online resource (XII, 416 pages) : 44 illustrations, 4 illustrations in color.
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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.
Antibody Preparation
Overview of Antibodies for Immunochemistry
to the Purification of Antibodies
Antibody Purification: Ammonium Sulfate Fractionation or Gel Filtration
Antibody Purification: Ion-Exchange Chromatography
Antibody Purification: Affinity Chromatography - Protein A and Protein G Sepharose
Conjugation of Fluorochromes to Antibodies
Biotinylation of Antibodies
Tissue Preparation for Light Microscopic Analysis
Cell Fixatives for Immunostaining
Permeabilization of Cell Membranes
Preparation of Frozen Sections for Analysis
Processing of Cytological Specimens
Processing of Tissue Culture Cells
Processing of Tissue Specimens
Heat-Induced Antigen Retrieval for Immunohistochemical Reactions in Routinely Processed Paraffin Sections
Bright Field Detection Systems
Fluorochromes: Properties and Characteristics
Direct Immunofluorescent Labeling of Cells
Fluorescence Labeling of Surface Antigens of Attached or Suspended Tissue-Culture Cells
Fluorescence Labeling of Intracellular Antigens of Attached or Suspended Tissue-Culture Cells
Fluorescent Visualization of Macromolecules in Drosophila Whole Mounts
Overview of Conventional Fluorescence Photomicrography
Overview of Confocal Microscopy
Overview of Laser Microbeam Applications as Related to Antibody Targeting
Immuno-Laser Capture Microdissection of Rat Brain Neurons for Real Time Quantitative PCR
Overview of Antigen Detection Through Enzymatic Activity
The Peroxidase-Antiperoxidase (PAP) Method and Other All-Immunologic Detection Methods
The Avidin-Biotin Complex (ABC) Method and Other Avidin-Biotin Binding Methods
Avidin-Biotin Labeling of Cellular Antigens in Cryostat-Sectioned Tissue
Multiple Antigen Immunostaining Procedures
Immunoenzymatic Quantitative Analysis of Antigens Expressed on the Cell Surface (Cell-ELISA)
Use of Immunogold with Silver Enhancement
Fluorescence-Activated Cell Sorter (FACS) Analyses
Overview of Flow Cytometry and Fluorescent Probes for Flow Cytometry
Tissue Disaggregation
Indirect Immunofluorescent Labeling of Viable Cells
Indirect Immunofluorescent Labeling of Fixed Cells
Fluorescent Labeling of DNA
Deparaffinization and Processing of Pathologic Material
Colloidal Gold Detection Systems for Electron Microscopic Analysis
Fixation and Embedding
Preparation of Colloidal Gold
Conjugation of Colloidal Gold to Proteins
Colloidal Gold/Streptavidin Methods
Pre-embedding Labeling Methods
Postembedding Labeling Methods
The Clinical Laboratory
The Clinical Immunohistochemistry Laboratory: Regulations and Troubleshooting Guidelines.
Oliver, Constance. editor.
Jamur, Maria Célia. editor.
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