During the last 30 years there has been a growing interest in cytokines as biological molecules able to regulate the most diverse functions in living org- isms, mainly at the level of cell-cell communication. Originally their definition was limited to the cells of the immune system (interleukins and lymphokines), but later that definition was extended to all cells, and their regulatory activity in such other processes as differentiation, apoptosis, angiogenesis, and wound he- ing has been now demonstrated. They comprise a group of small proteins (5-20 kDa) produced and released by cells in a tightly controlled fashion, active in the nano- or picomolar concentration range, and eliciting specific effects in nei- boring cells; therefore, their action is said to be autocrine, paracrine, or jux- crine. The latter property distinguishes them from hormones, which are produced by one tissue and are transported by the blood stream in order to act on a distant tissue. Chemokines are a subset of cytokines, but whether growth factors are included in the group is often a matter of discussion. The activity of several cytokines can be inhibited by other cytokines or by biological response modi- ers; therefore, the latter are sometimes called "anti-cytokines. " The biological response of a particular cell is usually the result of the sum of all interactions with cytokines present at a certain time and in a certain sequence in time-the "cytokine network. Contraportada 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.
- Libro Impreso
- Edición:
- Editorial: Vintage
- Autor: de Ley, Marc
