Cytokine Protocols (Methods in Molecular Biology Vol 249)

✍ Scribed by Marc De Ley

• Publisher: Humana Press
• Year: 2003
• Tongue: English
• Leaves: 239
• Edition: 1
• Category: Library

✦ Synopsis

A collection of biochemical, cellular, and molecular techniques for unraveling and quantifying the events occurring between the initial contact of a cytokine at the membrane receptor and the eventual activation of gene transcription. The techniques used include the generation of transfectants, the immunohistochemical detection of cytokines in tissue sections, and optimized staining for cytoplasmic detection. Highlights include RT-PCR of small amounts of mRNA, in situ hybridization, biosensor analysis, measurement of biological activities and standardization, immunohistochemical and single-cell detection, and receptor isolation, characterization, and crystallization.

✦ Table of Contents

Front......Page 1
1 Large-Scale Generation of Plasmids that Express Type I Interferon......Page 2
2 Identification of trans-Acting Factors by Electrophoretic Mobility Shift Assay......Page 8
3 Modulation of the Interferon-γ Signal by Transfection of Cells with an Antisense-RNA Expressing Vector......Page 22
4Competitive RT-PCR to Quantify Small Amounts of mRNA......Page 32
5 In Situ Hybridization for Cytokines in Human Tissue Biopsies......Page 42
6 Purification, Identification, and Synthesis of Chemokines......Page 48
7 Receptor Isolation and Characterization - From Protein to Gene......Page 66
8 Crystallization of Cytokine–Receptor Complexes......Page 82
9 Biosensor Analysis of Receptor–Ligand Interactions......Page 94
10 Analysis of SH2 Ligands and Identification of Sites of Interaction......Page 112
11 Assays for Antiviral Activity......Page 122
12 Assays for Cytotoxicity......Page 136
13 Assays for Chemotaxis......Page 154
14 In Vitro and In Vivo Assays for High-Molecular-Weight Pegylated Muteins of Granulocyte Colony-Stimulating Factor......Page 168
15 Development of a Mammalian Tet-On Expression Cell Line......Page 178
16 Immunohistochemical Detection of Cytokines in Human Tissue Sections......Page 194
17 Detection of Cytokine- and Chemokine-Expressing Cells at the Single Cell Level......Page 202
18 Detection of Cytokine Signal Transduction “Cross-Talk” in Leukocyte Activation......Page 220
19 Assay System for the Effect of Prostaglandin E2 in the Determination of Polarized Cytokine Production......Page 230