Cholesterol Homeostasis, Softcover reprint of the original 1st ed. 2017
Methods and Protocols
Methods in Molecular Biology Series, Vol. 1583

This volume provides state-of-the-art techniques for studying various aspects of cholesterol homeostasis, including its uptake, synthesis and efflux from the cell, as well as its trafficking within the cell.  Chapters also cover techniques for studying the regulation of cholesterol homeostasis at both the transcriptional and post-translational levels, as well as studying the membrane topology and structure of cholesterol-related proteins. 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.

Authoritative and practical, Cholesterol Homeostasis: Methods and Protocols aims to provide key techniques in tackling the investigation of cholesterol homeostasis.

An Overview of Cholesterol Homeostasis.- Hybrid In Silico/In Vitro Approaches for the Identification of Functional Cholesterol Binding Domains in Membrane Proteins.- Structural Stringency of Cholesterol for Membrane Protein Function utilizing Stereoisomers as Novel Tools: A Review .- Manipulating Cholesterol Status within Cells.- Assaying Low-Density-Lipoprotein (LDL) Uptake into Cells.- The use of L-sIDOL Transgenic Mice as a Murine Model to Study Hypercholesterolaemia and Atherosclerosis.- CRISPR/Cas9-mediated Generation of Niemann-Pick C1 Knockout Cell Line.- Quantitative Measurement of Cholesterol in Cell Population Using Flow Cytometry and Fluorescent Perfringolysin O*.- Transport Assays for Sterol-binding Proteins: Stopped-flow Fluorescence Methods for Investigating Intracellular Cholesterol Transport Mechanisms of NPC2 Protein.- Synthesis and Live-cell Imaging of Fluorescent Sterols for Analysis of Intracellular Cholesterol Transport.- Measurement of Cholesterol Transfer fromLysosome to Peroxisomes Using an In Vitro Reconstitution Assay.- Measurement of Mitochondrial Cholesterol Import using a Mitochondria-targeted CYP11A1 Fusion Construct.- Identifying Sterol Response Elements within Promoters of Genes.- Membrane Extraction of HMG CoA Reductase as Determined by Susceptibility of Lumenal Epitope to In Vitro Protease Digestion.- Determining the Topology of Membrane-bound Proteins using PEGylation.- Measuring Activity of Cholesterol Synthesis Enzymes Using Gas Chromatography/Mass Spectrometry.- Sterol Analysis by Quantitative Mass Spectrometry.- Measurement of Rates of Cholesterol and Fatty Acid Synthesis in vivo using Tritiated Water.- Methods for Monitoring ABCA1-dependent Sterol Release.- ABC-transporter Mediated Sterol Export from Cells using Radio-Labeled Sterols.- Measurement of Macrophage-specific In Vivo Reverse Cholesterol Transport in Mice.