Advanced Methods in Cellular Immunology

Published:
May 26, 2000 by CRC Press - 192 Pages
Author(s):
Rafael Fernandez-Botran, University of Louisville, Kentucky, USA; Vaclav Vetvicka, University of Louisville, Kentucky, USA

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$149.95
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ISBN 9780849321252
Cat# 2125
 

Features

  • Provides step-by-step procedures
  • Presents PCR techniques and covers both human and murine models
  • Devotes an entire chapter to techniques related to Apoptosis
  • Includes list of manufacturers of all chemicals and materials used in the procedures with addresses, phone numbers, and Web sites

    • Summary

    Immunologists as well investigators in other disciplines may often use protocols involving the isolation, cultures and characterization of different types of leukocytes. Advanced Methods in Cellular Immunology is a collection of techniques in an easy-to-use format.
    Each chapter provides readers with related program information, a step-by-step description of the methodology, alternative techniques, pertinent references, and information about commercial sources for materials and regents. In addition to leukocytes, the authors guide readers through the processes of cell culture as well as inflammation and autoimmunity in a variety of animal models.
    Covering topics such as PCR and Apoptosis, this book will serve a guide to commonly used procedures in cellular immunology while utilizing both human and murine models.

    Table of Contents

    SOURCES
    Preparation of Cell Suspensions
    Isolation of Cells
    Mitomycin C Treatment
    Preparation of Mycobacteria Suspension
    Preparation of Listeria Suspension
    ELISA
    Incorporation of TritiatedThymidine into DNA
    MTT Assay
    Cr Labeling
    ISOLATION AND CHARACTERIZATION OF NEUTROPHILS
    Percoll Sedimentation and Ficoll-Paque Centrifugation
    Magnetic Cell Sorting with Anti-CD15 Beads
    Functional Assessment
    ISOLATION AND CHARACTERIZATION OF EOSINOPHILS
    Discontinuous Density Percoll Gradients
    Magnetic Cell Sorting with Anti-CD16 Beads
    Functional Assessment
    ENRICHMENT OF HUMAN BASOPHILS
    MACROPHAGES
    Isolation of Macrophages
    Activation of Macrophages
    Production of Oxidative Metabolism Intermediates
    Phagocytosis
    Killing of Bacteria
    Killing of Bacteria - Colorimetric Evaluation
    Microbicidal Assay
    B LYMPHOCYTE CLONING
    Murine B Lymphocyte Cloning Assay
    Proliferation of Pre-B Cell Colonies in Methycellulose
    Clonal Assay for Murine Lymphohematopoietic Progenitors
    Clonal Assay for Fetal Cells
    Long-Term Bone Marrow Cultures
    CELL CULTIVATION IN-VITRO
    Tissue Culture Laboratory
    Aseptic Techniques
    Safety Concerns
    Substrate
    Basic Principles
    Primary Cultures
    Freezing of Cell Lines
    ANALYSIS OF CYTOKINE OR CYTOKINE RECEPTOR mRNA
    Introduction
    Total RNA Isolation
    Reverse Transcription-Polymerase Chain Reaction (RT-PCR)
    INTRACELLULAR CYTOKINE STAINING
    Cell Activation
    Staining
    CELL CYTOTOXICITY
    Preparation of Mouse Cytotoxic T Cells
    Whole Blood Assay
    DNA Fragmentation
    Apoptosis
    Quantitation of Apoptosis Using Fluorescent Dyes
    Quantitation of DNA Fragmentation
    Target Cell Detachment Assay
    Cytolysin Hemolytic Activity
    Isolation of Cytoplasmic Granules
    Esterase Assay
    Activation of Antibodies
    Activation by Phorbol Myristate Acetate and Ionophore
    MIXED LYMPHOCYTE REACTION
    Unidirectional MLR
    ANIMAL MODELS OF AUTOIMMUNE DISEASES
    Diabetes Mellitus
    Nonobese Mice
    Chemically Induced Diabetes
    Experimental Autoimmune Encephalomyelitis
    Rheumatoid Arthritis
    BUFFERS AND MEDIA
    COMMERCIAL SOURCES
    INDEX.

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