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Full Description
Featuring a diverse array of model organisms and scientific techniques, Sirtuins: Methods and Protocols collects detailed contributions from experts in the field addressing this vital family of genes. Opening with methods to generate sirtuin biology tools, the book continues by covering methods to identify sirtuin substrates, to measure sirtuin activity, and to study sirtuin biology. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and easy to use, Sirtuins: Methods and Protocols presents detailed protocols for sirtuin research that can be followed directly or modified to investigate new areas of sirtuin biology.
Contents
Introduction: Sirtuins in Aging and Diseases.- Sirtuins in Yeast: Phenotypes and Tools.- Studying the C. elegans Sirtuins.- Genetic and Biochemical Tools for Investigating Sirtuin Function in Drosophila melanogaster.- Generating Mammalian Sirtuin Tools for Protein-Interaction Analysis.- Mass Spectrometry-Based Detection of Protein Acetylation.- SILAC-Based Quantification of Sirt1-Responsive Lysine Acetylome.- Targeted Quantitation of Acetylated Lysine Peptides by Selected Reaction Monitoring Mass Spectrometry.- Identification of Deacetylase Substrates with the Biotin Switch Approach.- Assaying Chromatin Sirtuins.- Measurement of Sirtuin Enzyme Activity Using a Substrate-Agnostic Fluorometric Nicotinamide Assay.- Detecting Sirtuin-Catalyzed Deacylation Reactions Using 32P-Labeled NAD and Thin-Layer Chromatography.- Chemical Acetylation And Deacetylation.- Accurate Measurement of Nicotinamide Adenine Dinucleotide (NAD+) with High-Performance Liquid Chromatography.- In Vivo Measurement of the Acetylation State of Sirtuin Substrates as a Proxy for Sirtuin Activity.- Oxygen Flux Analysis to Understand the Biological Function of Sirtuins.- The Emerging Links Between Sirtuins and Autophagy.- Methods to Study the Role of Sirtuins in Genome Stability.- Circadian Measurements of Sirtuin Biology.- Utilizing Calorie Restriction to Evaluate the Role of Sirtuins in Healthspan and Lifespan of Mice.
