Histone Deacetylases Methods and Protocols

This volume provides different methodologies for all classes of histone deacetylases, which includes detailed procedures on Class I and II histone deacetylase inhibitors, SIRT inhibitors, and bromodomain inhibitors. Histone Deacetylases: Methods and Protocols is divided into four sections: Sections A and B describe methodologies used to detect the activity, function, or chromatin location of HDACs 1 through 11, with Section A discussing Class I and Section B discussing class II histone deacetylases; Section C focuses on the methodologies for cloning and characterizing the acetylation of SIRTs 1 through 7; and Section D describes methods related to histone deacetylase inhibitors. 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.

Thorough and cutting-edge, Histone Deacetylases: Methods and Protocols, is a valuable resource for investigators working on epigenetics, molecular biology, and genetics.

A Sensitive and Flexible Assay for Determining Histone Deacetylase 1 (HDAC1) Activity.- Detection of Sumo Modification of Endogenous Histone Deacetylase 2 (HDAC2) in Mammalian Cells.- Analysis of Epigenetic Regulation of Hypoxia-Induced Epithelial-Mesenchymal Transition in Cancer Cells by Quantitative Chromatin Immunoprecipitation of Histone Deacetylase 3 (HDAC3).- Molecular and Functional Characterization of Histone Deacetylase 4 (HDAC4).- Approaches for Studying the Subcellular Localization, Interactions, and Regulation of Histone Deacetylase 5 (HDAC5).- Analysis of Expression and Functions of Histone Deacetylase 6 (HDAC 6).- Analysis of Histone Deacetylase 7 (HDAC7) Alternative Splicing and Its Role in Embryonic Stem Cell Differentiation toward Smooth Muscle Lineage.- Large-Scale Overproduction and Purification of Recombinant Histone Deacetylase 8 (HDAC8) from the Human-Pathogenic Flatworm Schistosoma mansoni.- Visualization of Histone Deacetylase9 (HDAC9) Spatiotemporal Subcellular Localization in Primary Neuron Cultures.- Expression and Function of Histone Deacetylase 10 (HDAC10) in B Cell Malignancies.- Functional Analysis of Histone Deacetylase 11 (HDAC11).- Sirtuin1 (SIRT1) in the Acetylation of Downstream Target Proteins.- Protocols for Cloning, Expression, and Functional Analysis of Sirtuin2 (SIRT2).- Cloning and Characterization of Sirtuin3 (SIRT3).- Identification of Sirtuin4 (SIRT4) Protein Interactions: Uncovering Candidate Acyl-Modified Mitochondrial Substrates and Enzymatic Regulators.- Generation and Purification of Catalytically Active Recombinant Sirtuin5 (SIRT5) Protein.- Sirtuin6 (SIRT6) Activity Assays.- Molecular, Cellular, and Physiological Characterization of Sirtuin7 (SIRT7).- HDAC Inhibitors.- Assessment of the Anti-Proliferative Activity of a BET Bromodomain Inhibitor as Single Agent and in Combination in Non-Hodgkin Lymphoma Cell Lines.- Screening the Impact of Sirtuin Inhibitors on Inflammatory and Innate Immune Responses of Macrophages and in a Mouse Model of Endotoxic Shock.