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Neuroscientists, psychiatrists, pathologists, oncologists, surgeons, microbiologists, and other researchers and physicians from around the world present 36 contributions which address the major techniques in interleukin research methodologies currently available. The five sections address methods for assaying interleukin protein and mRNA, including the use of ELISA; the assays of interleukins in specific pathologies; the assaying of interleukins in different biological fluids relevant to disease; and newer methods in the study of interleukin signal transduction, analysis of polymorphisms in interleukin genes, and the use of cDNA arrays. Of likely interest to a wide array of scientific investigators including molecular and cell biologists and immunologists.Book News, Inc.

Book Info

Trinity College, Dublin, Ireland. Text offering biomedical researchers a collection of the major techniques used to assess the role of interleukins in disease, to facilitate diagnosis, and to speed the creation of new therapies. Features halftone illustrations. Designed as a laboratory manual for researchers.

Preface
Contributors
Pt. I General Methods for Assaying Interleukin Protein and mRNA 1
1 ELISAs and Interleukin Research 3
2 ELISPOT Technique for Assaying Interleukins 17
3 Measurement of Interleukins by ELISPOT Assay with Particular Application to Dual-Color Analysis (Stardust Assay) 29
4 Assaying Interleukins Intracellularly by FACS 41
5 Basic RT-PCR for Measurement of Cytokine Expression 49
6 Quantitative PCR for Measurement of Cytokine Expression 59
7 Single Cell RT-PCR for the Detection of Cytokine mRNA 77
8 Quantitation of Cytokine mRNA by Flash-Type Bioluminescence 83
9 Measurement of Interleukin mRNA by Northern Blotting Using a Nonradioactive Detection Method 99
10 Quantification of Mouse IL-6 and TNF-[alpha] mRNA Using Xplore Assays 111
11 Measurement of Cytokine and Chemokine mRNA Using Nonisotopic Multiprobe RNase Protection Assay 125
12 Isolation of Total and Bioactive Interleukins by Immunoaffinity-Receptor Affinity Chromatography 145
13 Detection of Cytokines by Immunohistochemistry 157
14 Whole Blood Assays and the Influence of Circadian Rhythmicity on Human Cytokine Measurement 163
Pt. II Illustration of Methodologies Using Individual Interleukins 175
15 Assaying Interleukin-8 177
16 Measurement of Interleukin-2 by Bioassay 185
17 Assaying Interleukin-15 193
Pt. III Assaying Interleukins in Particular Pathologies 205
18 Assaying Interleukin-6 in Breast Cancer 207
19 Cytokines, Stress, and Depressive Illness 217
20 Measurement of Interleukins in Cutaneous Disorders: Competitive RT-PCR for Determination of IL-10 mRNA Expression in Psoriasis 239
21 Interleukins in Graves' Disease 251
22 Evaluation of Cytokines in Migraine Patients by ELISA 263
23 Assaying Interleukins in Plasma in the Course of Acute Myocardial Infarction 273
Pt. IV Assaying Interleukins in Particular Biological Fluids 283
24 Measurement of Cytokines in Peritoneal Fluids 285
25 Measurement of Cytokines in Seminal Plasma 293
26 Measurement of Cytokines in Induced Sputum: Application to the Investigation of Asthma and COPD 305
27 Cytokines in Blister Fluids 313
28 Measurement of Cytokines in Synovial Fluid 327
29 Measurement of Cytokines in Cerebrospinal Fluid 337
30 Detection of Cytokines in Tears 345
Pt. V Newer Techniques: Interleukin Signal Transduction, Recombinant Interleukin Production, Interleukin Gene Polymorphisms, and the Use of cDNA Arrays 355
31 Measurement of Phosphoinositide 3-Kinase Activation by Interleukins 357
32 Analysis of Interleukin-2 Signaling Using Affinity Precipitations and Polyacrylamide Gel Electrophoresis 369
33 Applications for Green Fluorescent Protein in Cell Signaling 385
34 Use of Pichia pastoris for Production of Recombinant Cytokines 393
35 Analysis of Single Nucleotide Polymorphisms in the Interleukin-1B Gene Using 5' Nuclease Assays 409
36 Measurement of Cytokine and Chemokine Gene Expression Patterns Using cDNA Array 419
Index 439