Mammalian cells have evolved a complex multicomponent machinery that enables them to sense and respond to a wide variety of potentially toxic agents present in their environment. These stress responses are often associated with an increased cellular capacity to tolerate normally lethal levels of an insult. The realization that the mammalian stress response may be intimately linked with many human diseases, including rheumatoid arthritis, ischemia, fever, infection, and cancer, has led to an explosion of interest in this research area. Stress Response: Methods and Protocols brings together a diverse array of practical methodologies that may be employed to address various aspects of the response of mammalian cells to environmental stress. The p- tocols are carefully described by authors who have both devised and succe- fully employed them, and they represent a mixture not only of well-established techniques, but also new technologies at the leading edge of research. The areas covered include the detection and assay of stress-induced damage, the acti- tion of signal transduction pathways, stress-inducible gene expression, and stress protein function. Although no volume of this size can be comprehensive and the topics covered reflect a personal choice, it is hoped that it will prove of subst- tial interest and use to a wide range of research workers in the field.

Klappentext

In is now understood that the response of mammalian cells to a wide variety of potentially toxic agents may be intimately linked with many human diseases, including rheumatoid arthritis, ischemia, fever, infection, and cancer. In Stress Response: Methods and Protocols, Stephen Keyse has assembled a diverse collection of readily reproducible methods devoted to the study of these varied and powerful responses. Written by leading researchers expert in the techniques they describe, these detailed methods cover the detection and assay of stress-induced damage, the activation of a wide range of signal transduction pathways by cellular stress, stress-induced gene expression, and stress protein function. To ensure experimental success, step-by-step guidance is provided for each method, along with details of reagents, equipment, and other requirements. The methods include both well-established techniques and new technologies at the leading edge of research.
Wide ranging and highly practical, Stress Response: Methods and Protocols provides a gold-standard bench manual for today's basic and clinical scientists working to understand how cells and tissues respond during physiological stress and in human disease

Zusammenfassung
Stress Response: Methods and Protocols brings together a diverse array of practical methodologies that may be employed to address various aspects of the response of mammalian cells to environmental stress.

Inhalt
Detection and Assay of Stress-Induced Damage.- Identifying and Counting Protein Modifications Triggered by Nitrosative Stress.- Determination of Carbonyl Groups in Oxidized Proteins.- Quantitation of 4-Hydroxynonenal Protein Adducts.- Detection of Oxidative Stress in Lymphocytes Using Dichlorodihydrofluorescein Diacetate.- The Measurement of Protein Degradation in Response to Oxidative Stress.- The Activation of Signal Transduction by Cellular Stress.- Analysis of the Role of the AMP-Activated Protein Kinase in the Response to Cellular Stress.- Detection and Activation of Stress-Responsive Tyrosine Kinases.- Detection of DNA-Dependent Protein Kinase in Extracts from Human and Rodent Cells.- Expression and Assay of Recombinant ATM.- Detection and Purification of a Multiprotein Kinase Complex from Mammalian Cells.- Methods to Assay Stress-Activated Protein Kinases.- Monitoring the Activation of Stress-Activated Protein Kinases Using GAL4 Fusion Transactivators.- Use of Kinase Inhibitors to Dissect Signaling Pathways.- The Development and Use of Phospho-Specific Antibodies to Study Protein Phosphorylation.- Peptide Assay of Protein Kinases and Use of Variant Peptides to Determine Recognition Motifs.- The Analysis of Stress-Induced Gene Expression.- Assaying NF-KB and AP-1 DNA-Binding and Transcriptional Activity.- Analysis of the Mammalian Heat-Shock Response.- Approaches to Define the Involvement of Reactive Oxygen Species and Iron in Ultraviolet-A Inducible Gene Expression.- The Human Immunodeficiency Virus LTR-Promoter Region as a Reporter of Stress-Induced Gene Expression.- Sage: The Serial Analysis of Gene Expression.- Analysis of Differential Gene Expression Using the SABRE Enrichment Protocol.- UVB-Regulated Gene Expression in Human Keratinocytes.- Analysis of Stress Protein Function.- Heme Oxygenase Activity Current Methods and Applications.- Analysis of Molecular Chaperone Activities Using In Vitro and In Vivo Approaches.- Analysis of Chaperone Properties of Small Hsp'.- Analysis of Small Hsp Phosphorylation.- Analysis of Multisite Phosphorylation of the p53 Tumor-Suppressor Protein byTryptic Phosphopeptide Mappin.- Development of Physiological Models to Study Stress Protein Responses.