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Immunocytochemical Methods and Protocols, 2nd Edition

  •  ISBN-10 0-89603-570-0

  •  ISBN-13 978-0-89603-570-6

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publication info   L.C. Javois, ed., Humana Press, 1999, 465 pp., hard cover
mfr. no.   Humana

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General description

This 2nd edition revises and updates the previous collection of step-by-step immunocytochemical methods now used in many biological and biomedical research programs. The methods are designed for researchers and clinicians that wish to visualize molecules in plant or animal embryos, tissue sections, cells, or organelles. In addition to cutting-edge protocols for purifying and preparing antibodies, light microscopic analysis, confocal microscopy, FACS, and electron microscopy are also covered.

Table of Contents

Part I. Antibody Preparation.
Overview of Antibody Use in Immunocytochemistry.
Purification of Antibodies Using Ammonium Sulfate Fractionation or Gel Filtration,
Purification of Antibodies Using Ion-Exchange Chromatography,
Purification of Antibodies Using Affinity Chromatography.
Purification of Antibodies Using Protein A-Sepharose® and FPLC.
Conjugation of Fluorochromes to Antibodies,
Biotinylation of Antibodies.
Part II. Tissue Preparation for light Microscopic Analyses.
Overview of Cell Fixatives and Cell Membrane Permeants,
Preparation of Frozen Sections for Analysis,
Processing of Cytological Specimens.
Processing of Tissue Culture Cells,
Processing of Tissue Specimens,
Antigen Retrieval for Immunohistochemical Reactions in Routinely Processed Paraffin Sections,
Part III. Light Microscopic Detection Systems.
Overview of Fluorochromes,
Direct Immunofluorescent Labeling of Cells.
Fluorescence Labeling of Surface Antigens on Attached or Suspended Tissue-Culture Cells.
Fluorescence Labeling of Intracellular Antigens in Attached or Suspended Tissue-Culture Cells.
Fluorescent Labeling of Surface or Intracellular Antigens in Whole-Mounts.
TUNEL Assay for Apoptotic Cells,
Overview of Fluorescence Analysis with the Confocal Microscope.
Overview of Laser Microbeam Applications as Related to Antibody Targeting,
Overview of Fluorescence Photomicography.
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,
ELISA on Attachment Dependent or Suspension Grown Cells,
Use of Immunogold with Silver Enhancement,
Part IV. Fluorescence Activated Cell Sorter (FACS) Analyses.
Overview of Flow Cytometry and Fluorescent Probes for 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, Assay for Phagocytosis.
Assay for Filamentous Actin,
Assay for Chemoattractant Binding,
Assay for Oxidative Metabolism,
Part V. Colloidal Gold Detection Systems For Electron Microscopic Analyses. Fixation and Embedding,
Preparation of Colloidal Gold, Conjugation of Colloidal Gold to Proteins.
Colloidal Gold/Streptavidin Methods, Pre-Embedding Labeling Methods, Postembedding Labeling Methods,
Part VI. Applications For Nucleic Acid Localization By In Situ Hybridization. Overview of In Situ Hybridization,
In Situ Hybridization to Human Chromosomes of an Alkaline Phosphatase-Labeled Centromeric Probe,
Fluorescence In Situ Hybridization Using Whole Chromosome Library Probes,
In Situ RT-PCR and Hybridization Techniques,
Part VII. Special Applications For the Clinical Laboratory.
Overview of the Clinical Immunohistochemistry Laboratory: Regulations and Troubleshooting Guidelines,
Overview of

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