Amyloid Proteins Methods and Protocols

This third edition volume expands on the previous editions with many new chapters that cover the latest techniques and topics that were not addressed in the previous volumes. The chapters in this book are divided into three parts: Part One covers in vitro assays that focus on a variety  of amyloids and how to study these peptides and proteins. Part Two describes cell culture models and assays, and Part Three explores methods on how to extract amyloid from tissue, its detection, and its characterization in vitro or in vivo. 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.

Cutting-edge and authoritative, Amyloid Proteins: Methods and Protocols, Third Edition is a valuable resource for both students and scientists who are new to the field, as well as experienced researchers who would like to progress their research with the latest available techniques.

Preparation of Pure Populations of Amyloid-ß Protein Oligomers of Defined Size.- Preparation of Well-Defined and Stable ß-Barrel Pore-Forming Amyloid-ß42 Oligomers.- Unveiling Brain Aß Heterogeneity through Targeted Proteomic Analysis.- Preparation of a-Synuclein Amyloid Assemblies for Toxicity Experiments.- Generation and Characterization of Stable a-Synuclein Oligomers.- In Vitro Analysis of a-Synuclein Amyloid Formation and Cross-Reactivity.- Preparation of Tau Oligomers after Protein Extraction from Bacteria and Brain Cortices.- Purification and Characterization of Low-n Tau Oligomers.- Preparation and Characterization of Tau Oligomer Strains.- Purification and Fibrillation of Recombinant Human Amyloid-ß, Prion Protein and Tau under Native Conditions.- Preparation of Amyloidogenic  Aggregates from EF-Hand ß –parvalbumin and S100 Proteins.- Kinetic Analysis of Amyloid Formation.- Mapping Amyloid Regions in Gad m 1with Peptide Arrays.- Non-Invasive Structural Analysis of Intermediate Species during Fibrillation: An Application of Small-Angle X-Ray Scattering.- Analysis of Covalent Modifications of Amyloidogenic Proteins using Two-Dimensional Electrophoresis: Prion Protein and Its Sialylation.- Amplification and Detection of Minuscule Amounts of Misfolded Prion Protein by Using The Real-Time Quaking-Induced-Conversion.- Bacterial Amyloids.- Addressing Intracellular Amyloidosis in Bacteria with RepA-WH1, a Prion-Like Protein.- Study of Amyloids Using Yeast.- Neurotoxic Ca2+ Signaling Induced by Amyloid-ß Oligomers in Aged Hippocampal Neurons In Vitro.- Preparation and Culturing of Human Primary Vascular Cells.- Live Imaging of Pathological Tau Protein and Tau Antibodies in a Human Neuron-Like Cellular Model.- Effects of Amyloid-ß Peptide on the Biology of Human Neural Stem Cells.- Development of Mouse Monoclonal Antibodies against Human Amyloid Fibril Proteins for Diagnostic and Research Purposes.- Identification and Characterization of Amyloid-ß Accumulation in Synaptic Mitochondria.- Biochemical Properties of Pathology-Related Tau Species in Tauopathy Brains: An Extraction Protocol for Tau Oligomers and Aggregates.- Tau Assembly into Filaments.- Quantitative Metabolomics in Alzheimer´s Disease: Technical Considerations for Improved Reproducibility.- Detecting Circulating MicroRNAs as Biomarkers in Alzheimer’s Disease.- Luminescent Conjugated Oligothiophene Probe Applications for Fluorescence Imaging of Pure Amyloid Fibrils and Protein Aggregates in Tissues.- Characterization of Amyloid-ß Plaques and Autofluorescent Lipofuscin Aggregates in Alzheimer´s Disease Brain: A Confocal Microscopy Approach.- In Vivo Imaging of Tauopathy in Mice.- In Vivo Evaluation of Neuronal Transport in Murine Models of Neurodegeneration using Manganese-Enhanced MRI.