Because cytokines regulate many different bodily functions in
living organisms, mainly at the level of cell-cell communications,
there is great interest in understanding their mode of action. In
Cytokine Protocols, established researchers, physicians, and
clinicians present their best 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. Described in
step-by-step detail to ensure successful experimental results,
these protocols cover the large-scale generation and purification
of plasmid DNA, the identification of DNA-protein interactions via
the gel mobility shift assay, RNA-level phenomena, and the
isolation and characterization of cytokines, cytokine-related
proteins, and their interactions. 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. Each protocol includes a background
introduction, equipment and reagent lists, tips on troubleshooting
and avoiding pitfalls, and, where needed, a discussion of the
interpretation of results. Comprehensive and highly practical,
Cytokine Protocols offers novice researchers a concise introduction
to the principles of cytokine research, and experienced
investigators all the advanced techniques and time-saving tips
needed to ensure productive research.
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