Amyloid Proteins / Edition 1

by Einar M. Sigurdsson

Overview

A proven collection of readily reproducible techniques for studying amyloid proteins and their involvement in the etiology, pathogenesis, diagnosis, and therapy of amyloid diseases. The contributors provide methods for the preparation of amyloid and its precursors (oligomers and protofibrils), in vitro assays and analytical techniques for their study, and cell culture models and assays for the production of amyloid proteins. Additional chapters present readily reproducible techniques for amyloid extraction from tissue, its detection in vitro and in vivo, as well as nontransgenic methods for developing amyloid mouse models. The prools follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.

Product Details

ISBN-13:	9781617375026
Publisher:	Springer-Verlag New York, LLC
Publication date:	11/19/2010
Series:	Methods in Molecular Biology , #299
Edition description:	Softcover reprint of hardcover 1st ed. 2005
Pages:	408
Product dimensions:	5.98(w) x 9.02(h) x 0.36(d)

Table of Contents

In Vitro Assays.- Preparation of Aggregate-Free,Low Molecular Weight Amyloid? for Assembly and Toxicity Assays.- Determination of Peptide Oligomerization State Using Rapid Phohemical Crosslinking.- In Vitro Preparation of Prefibrillar Intermediates of Amyloid-? and— Synuclein.- Purification of Recombinant Tau Protein and Preparation of Alzheimer-Paired Helical Filaments In Vitro.- Cyclic Amplification of Protein Misfolding and Aggregation.- X-Ray Diffraction Studies of Amyloid Structure.- Molecular Electron Microscopy Approaches to Elucidating the Mechanisms of Protein Fibrillogenesis.- Time -Lapse Atomic Force Microscopy in the Characterization of Amyloid-Like Fibril Assembly and Oligomeric Intermediates.- Fourier Transform Infrared and Circular Dichroism Spectroscopies for Amyloid Studies.- Quasielastic Light Scattering for Protein Assembly Studies.- Intrinsic Fluorescent Detection of Tau Conformation and Aggregation.- Quantitative Measurement of Fibrillogenesis by Mass Spectrometry.- Cell Culture Assays.- Isolation and Culturing of Human Vascular Smooth Muscle Cells.- Murine Cerebrovascular Cells as a Cell Culture Model for Cerebral Amyloid Angiopathy.- Purification of Human Wild-Type or Variant Cystatin C From Conditioned Media of Transfected Cells.- Prion Propagation in Cell Culture.- In Vivo-Related Assays.- Preparation and Propagation of Amyloid-Enhancing Factor.- Purification of Amyloid Protein AA Subspecies From Amyloid-Rich Human Tissues.- Purification of Transthyretin and Transthyretin Fragments From Amyloid-Rich Human Tissues.- Extraction and Chemical Characterization of Tissue-Deposited Proteins From Minute Diagnostic Biopsy Specimens.- Tissue Processing Prior to Protein Analysis and Amyloid-ß Quantitation.- ELISA Method for Measurement ofAmyloid-ß Levels.- Histological Staining of Amyloid-ß in Mouse Brains.- The Mouse Model for Scrapie.- Radiolabeling of Amyloid-ß Peptides.- In Vivo Imaging of Amyloid-ß Deposits in Mouse Brain With Multiphoton Microscopy.- Magnetic Resonance Imaging of Amyloid Plaques in Transgenic Mice.

Editorial Reviews

2 Stars from Doody

Reviewer: Eugene A Davidson, PhD (Georgetown University School of Medicine)
Description: This is a compilation of protocols for working with protein aggregates associated with disease with a major focus on A-beta.
Purpose: The goal of this book is to provide a set of protocols of use to investigators working with amyloidogenic proteins. Given the complexity of the field, this is a generally worthwhile objective. Success is limited (incomplete coverage, lack of a unifying overview).
Audience: The target audience includes advanced students and fellows as well as senior investigators. The editor has recruited a diverse group of contributors working in the field.
Features: The increasing awareness of "folding diseases" has stimulated considerable research into the mechanisms of protein aggregation in these conditions. This book provides a series of protocols for working with several representatives of this class. It is divided into three sections, all of which define assays for measuring various properties of aggregate precursors or the aggregates themselves (in vitro, in vivo and cell culture). The major focus is on those proteins associated with Alzheimer's disease - A-beta and tau. The in vitro techniques reviewed are mainly physical methods (light scattering, AFM, FTIR) but include discussion of protein cross-linking and preparation of materials. Protocols are also provided for radiolabeling and purification of specific proteins. Each protocol has a short bibliography as well as detailed laboratory directions. The animal models are mainly those using mice. However, problems associated with these (very long incubation time for prion transmission, for example) are not discussed. In addition, regulatory concerns should be addressed. The article on propagation of prions in cell culture, written by a European investigator, does not address the restrictions in force in the U.S. for working with such materials. The book would have benefited from tighter editing and a broad overview summary. The literature in this field has expanded exponentially in recent years. Thus, this distillate will have value for interested investigators.
Assessment: As with many books of compiled protocols, this one needs better editing and some unifying overview. Investigators contributing protocols (often derived from their own work) rarely do a suitable job of identifying pitfalls.

Doody's Review Service