Cytokine protocols / edited by Marc de Ley.

Format:

Book

Contributor:

Ley, Marc de.

Series:

Methods in molecular biology (Clifton, N.J.) ; v. 249.

Methods in molecular biology, 1064-3745 ; 249

Language:

English

Subjects (All):

Cytokines--Laboratory manuals.

Cytokines.

Immunologic Techniques.

Medical Subjects:

Cytokines.

Immunologic Techniques.

Genre:

Laboratory manuals.

Physical Description:

xiii, 240 pages : illustrations ; 24 cm.

Place of Publication:

Totowa, N.J. : Humana Press, [2004]

Summary:

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.

Contents:

1 Large-Scale Generation of Plasmids that Express Type I Interferon / Peter Harle, Daniel J. J. Carr 1

2 Identification of trans-Acting Factors by Electrophoretic Mobility Shift Assay / Rebecca J. Chan, Min You, Gen-Sheng Feng 7

3 Modulation of the Interferon-[gamma] Signal by Transfection of Cells with an Antisense-RNA Expressing Vector / Leen Delrue, Marc De Ley 21

4 Competitive RT-PCR to Quantify Small Amounts of mRNA / Gabriela Aust 31

5 In Situ Hybridization for Cytokines in Human Tissue Biopsies / Emily Mathey, John Pollard, Patricia Armati 41

6 Purification, Identification, and Synthesis of Chemokines / Patricia Menten, Anja Wuyts, Jo Van Damme 47

7 Receptor Isolation and Characterization: From Protein to Gene / Daniela Novick, Menachem Rubinstein 65

8 Crystallization of Cytokine-Receptor Complexes / William J. Cook, Mark R. Walter 81

9 Biosensor Analysis of Receptor-Ligand Interactions / Sheng-Jiun Wu, Irwin Chaiken 93

10 Analysis of SH2 Ligands and Identification of Sites of Interaction / Manuel Baca 111

11 Assays for Antiviral Activity / Anthony Meager 121

12 Assays for Cytotoxicity / Anthony Meager 135

13 Assays for Chemotaxis / Anja Wuyts, Patricia Menten, Nancy Van Osselaer, Jo Van Damme 153

14 In Vitro and In Vivo Assays for High-Molecular-Weight Pegylated Muteins of Granulocyte Colony-Stimulating Factor / James F. Eliason, Nadine Tare, Sharon Bowen, Tomoaki Inoue, Ikuro Horii 167

15 Development of a Mammalian Tet-On Expression Cell Line: Glucosylceramide Synthase Regulates TNF-[alpha]-Induced Apoptosis / Yong-Yu Liu, Myles C. Cabot 177

16 Immunohistochemical Detection of Cytokines in Human Tissue Sections / Sandra Hulshof, Danny F. Dukers, Wim Vos, Corline J. A. De Groot 193

17 Detection of Cytokine- and Chemokine-Expressing Cells at the Single Cell Level / Karin Lore, Jan Andersson 201

18 Detection of Cytokine Signal Transduction "Cross-Talk" in Leukocyte Activation / Howard R. Petty 219

19 Assay System for the Effect of Prostaglandin E[subscript 2] in the Determination of Polarized Cytokine Production / Etsushi Kuroda, Uki Yamashita 229.

Notes:

Includes bibliographical references and index.

ISBN:

089603948X

1592596673

OCLC:

52086158