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T Cell Protocols

  •  ISBN-10 0-89603-810-6

  •  ISBN-13 978-0-89603-810-3

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publication info   K.P. Kearse, ed., Humana Press, 2000, 384 pp., hard cover
mfr. no.   Humana

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General description

A compilation of step-by-step techniques for the study of T cell developmental biology and their subsequent activation. It includes treatment of both fundamental and novel experimental protocols for the analysis of the role T cells play in immune function and in the prevention and cure of human diseases.

Table of Contents

PART I. Insights into T-Cell Development Using Transgenic and Knockout Mice,
PART II. T Cell Development Isolation and Characterization of Murine Early Intrathymic Precursor Populations,
Differentiation of Mouse Thymocytes in Fetal Thymus Organ Culture,
Purification of Immature CD4+CD8+ Thymocytes by Panning with Anti-CD8 Antibody,
Transfection and Transcription of Genes in Developing Thymocytes,
Analysis of CD4/CD8 Lineage Commitment by Pronase Treatment and Reexpression Assay,
Intrathymic Delivery of MHC Genes Using Recombinant Adenoviruses,
Quantitative Analysis of the Usage of Human T-Cell Receptor a and b Chain Variable Regions by Reverse Dot Blot Hybridization,
Flow Cytometric Detection of Cytoplasmic and Surface CD3-e Expression in Developing T Cells,
Assessing Apoptosis of Developing T Cells by Flow Cytometry,
Isolation of T-Cell Subsets by Magnetic Cell Sorting (MACS),
Isolation of T-Cell Antigens by Retrovirus-Mediated Expression Cloning,
Flow Cytometric Analysis of Murine T Lymphocytes: A Practical Guide,
Production and Characterization of Immature Murine T-Lymphoma Cell Lines,
Production and Characterization of T-Cell Hybridomas,
PART III. Overview: Something Happens.
A Brief History of TCR Signal Transduction,
PART IV. T Cell Activation. Measurement of Phosphatidylinositol (PI) Hydrolysis in Activated T Lymphocytes,
Fluorescence Polarization as an Early Measure of T-Lymphocyte Stimulation,
Measurement of Lymphoproliferation at the Single-Cell Level by Flow Cytometry,
Digital Image Analysis of Lymphocyte Activation, Induction of T-Cell Signaling by Immobilized Integrin Ligands,
Cytotoxic T-Cell Adherence Assay (CAA),
Detecting Ubiquitinated T-Cell Antigen Receptor Subunits by Immunoblotting,
Determination of CD45 Tyrosine Phosphatase Activity in T Lymphocytes,
Measurement of Protein Tyrosine Phosphorylation in T-Cell Subsets by Flow Cytometry,
Biochemical Analysis of Activated T Lymphocytes: Protein Phosphorylation and Ras, ERK, and JNK Activation,
Activation of Heterotrimeric GTP-Binding Proteins Upon TCR/CD3 Engagement,
Generation of Human T Helper 1 and T Helper 2 Subsets from Peripheral Blood-Derived Naive CD4+ T Cells,
Transcriptional Expression Analysis of T-Cell Activation by Multiplex Messenger Assay (MMA),
Index

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