De Ley M. Cytokine Protocols

Humana Press Inc., Totowa, NJ. - 2003. -248 p. Methods in Molecular Biology, vol. 249A 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.Large-Scale Generation of Plasmids that Express Type I Interferon
Identification of trans-Acting Factors by Electrophoretic Mobility Shift Assay
Modulation of the Interferon-[gamma] Signal by Transfection of Cells with an Antisense-RNA Expressing Vector
Competitive RT-PCR to Quantify Small Amounts of mRNA
In Situ Hybridization for Cytokines in Human Tissue Biopsies
Purification, Identification, and Synthesis of Chemokines
Receptor Isolation and Characterization: From Protein to Gene
Crystallization of Cytokine-Receptor Complexes
Biosensor Analysis of Receptor-Ligand Interactions
Analysis of SH2 Ligands and Identification of Sites of Interaction
Assays for Antiviral Activity
Assays for Cytotoxicity
Assays for Chemotaxis
In Vitro and In Vivo Assays for High-Molecular-Weight Pegylated Muteins of Granulocyte Colony-Stimulating Factor
Development of a Mammalian Tet-On Expression Cell Line: Glucosylceramide Synthase Regulates TNF-[alpha]-Induced Apoptosis
Immunohistochemical Detection of Cytokines in Human Tissue Sections
Detection of Cytokine- and Chemokine-Expressing Cells at the Single Cell Level
Detection of Cytokine Signal Transduction "Cross-Talk" in Leukocyte Activation
Assay System for the Effect of Prostaglandin E[subscript 2] in the Determination of Polarized Cytokine Production