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 protocols 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.

Part I. In Vitro Assays
Preparation of Aggregate-free, Low Molecular Weight Ab for Assembly and Toxicity Assays
Gal Bitan and David B. Teplow
Determination of Peptide Oligomerization State using Rapid Photochemical Cross-Linking
Sabrina S. Vollers, David B. Teplow, and Gal Bitan
In Vitro Preparation of Prefibrillar Intermediates of Amyloid-b (Ab) and a-Synuclein
Hilal A. Lashuel and Dolors Grillo-Bosch
Purification of Recombinant Tau Protein and Preparation of Alzheimer Paired Helical Filaments in Vitro
Stefan Barghorn, Jacek Biernat, and Eckhard Mandelkow
Cyclic Amplification of Protein Misfolding and Aggregation
Paula Saa, Joaquin Castilla, and Claudio Soto
X-Ray Diffraction Studies of Amyloid Structure
O Sumner Makin and Louise C Serpell
Molecular Electron Microscopy Approaches to Elucidating the Mechanisms of Protein Fibrillogenesis
Hilal A. Lashuel and Joseph Wall
Time-Lapse Atomic Force Microscopy in the Characterisation of Amyloid-Like Fibril Assembly and Oligomeric Intermediates
Claire Goldsbury and Janelle Green
FTIR and CD Spectroscopies for Amyloid Studies
Miguel Calero and Maria Gasset
Quasielastic Light Scattering for Protein Assembly Studies
Aleksey Lomakin, David B. Teplow, and George B. Benede
Intrinsic Fluorescent Detection of Tau Conformation and Aggregation
Martin von Bergen, Li Li, and Eckhard Mandelkow
Quantitative Measurement of Fibrillogenesis by Mass Spectrometry
Andrew D. Miranker
Part II. Cell Culture Assays
Isolation and Culturing of Human Vascular Smooth Muscle Cells
Finnbogi R. Thormodsson and Ingvar H. Olafsson
Murine Cerebrovascular Cells as a Cell Culture Model for Cerebral Amyloid Angiopathy: Isolation of Smooth Muscle and Endothelial Cells from Mouse Brain
Sonia S. Jung and Efrat Levy
Purification of Human Wild Type or Variant Cystatin C from Conditioned Media of Transfected Cells
Frances Prelli, Monika Pawlik, Blas Frangione, and Efrat Levy
Prion Propagation in Cell Culture
Sylvain Lehmann
Part III. In Vivo Related Assays
Preparation and Propagation of Amyloid Enhancing Factor
Robert Kisilevsky
Purification of Protein AA Subspecies from Amyloid-Rich Human Tissues
Gunilla T. Westermark and Per Westermark
Purification of Transthyretin and Transthyretin Fragments from Amyloid-Rich Human Tissues
Per Westermark and Gunilla T. Westermark
Extraction and Chemical Characterization of Tissue Deposited Proteins from Minute Diagnostic Biopsy Specimens
Fernando Goni and Gloria Gallo
Tissue Extraction Prior to Protein Analysis and Ab Quantitation
Stephen D. Schmidt, Ying Jiang, Ralph A. Nixon, and Paul M. Mathews
ELISA Method for Measurement of Ab Levels
Stephen D. Schmidt, Ralph A. Nixon, and Paul M. Mathews
Histological Staining of Amyloid-b in Mouse Brains
Einar M. Sigurdsson
The Mouse Model for Scrapie: Inoculation, Clinical Scoring, Histopathological and Immunohistopathological Techniques
Harry C. Meeker, Xuemin Ye, and Richard I. Carp
Radiolabeling of Amyloid-b Peptides
Miguel Calero and Jorge Ghiso
In vivo Imaging of Amyloid-b Deposits in Mouse Brain with Multiphoton Microscopy
Jesse Skoch, Gregory A. Hickey, Stephen T. Kajdasz, Bradley T. Hyman, and Brian J. Bacskai
Magnetic Resonance Imaging of Amyloid Plaques in Transgenic Mice
Youssef Zaim Wadghiri, Einar M. Sigurdsson, Thomas Wisniewski, and
Daniel H. Turnbull