Description
This volume details protocols on prion disease from multiple disciplines and highlights the contribution each discipline has made to the understanding of prion disease. 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, Prions: Methods and Protocols aims to ensure successful results in the further study of this vital field. PART I: Methods for Biochemical and Biophysical Analysis of Prion Disease 1. Purification and Fibrillation of Full-length Recombinant PrP Natallia Makarava, Regina Savtchenko, and Ilia V. Baskakov 2. Method for Folding of Recombinant Prion Protein to Soluble b-sheet Secondary Structure Laura J. Ellett 3. Analysis of Prion Protein Conformation using Circular Dichroism Spectroscopy Laura J Ellett and Vanessa Johanssen 4. Analysis of Prion Protein Structure using Nuclear Magnetic Resonance Spectroscopy Ivana Biljan, Gregor Ilc, and Janez Plavec 5. Immunodetection of PrPSc using Western Immunoblotting Techniques Gerald S. Baron and Gregory J. Raymond 6. Analysis of miRNA Signatures in Neurodegenerative Prion Disease Shayne A. Bellingham and Andrew F. Hill PART II: Methods for Cellular Analysis Of Prion Disease 7. Cell Biology Approaches to Studying Prion Diseases Suzette A. Priola 8. Expression of Heterologous PrP and Prion Propagation in RK13 Cells Zaira E. Arellano-Anaya, Alvina Huor, Pascal Leblanc, Olivier Androletti, and Didier Vilette 9. Generation of Infectious Prions and Detection with the Prion Infected Cell Assay Laura J Vella, Bradley Coleman, and Andrew F Hill 10. Analysis of Cellular Prion Protein Endoproteolytic Processing Victoria Lewis 11. Cellular Analysis Adult Neural Stem Cells for Investigating Prion Biology Cathryn L Haigh 12. Neurotoxicity of Prion Peptides on Cultured Cerebellar Neurons Giuseppe D. Ciccotosto, Metta Jana, and Roberto Cappai PART III: Methods for Cell Free Propagation of Prions 13. Methods of Protein Misfolding Cyclic Amplification Natallia Makarava, Regina Savtchenko, and Ilia V. Baskakov 14. RT-QuIC Assays for Prion Disease Detection and Diagnostics Christina D. Orr, Bradley R. Groveman, Andrew G. Hughson, Matteo Manca, Lynne D. Raymond, Gregory J. Raymond, Katrina J. Campbell, Kelsie J. Anson, Allison Kraus, and Byron Caughey 15. A Quick Method to Evaluate the Effect of the Amino Acid Sequence in the Misfolding Proneness of the Prion Protein Natalia Fernndez-Borges, Hasier Eraa, Saioa Rodrguez-Elezgarai, Chafik Harrathi, Vanesa Venegas, and Joaqun Castilla PART IV: In vivo Investigation of Prion Disease 16. Insights into Mechanisms of Transmission and Pathogenesis from Transgenic Mouse Models of Prion Diseases Julie A. Moreno and Glenn C. Telling 17. In vivo Near Infrared Imaging of Neurodegeneration Victoria A Lawson, Carolin Tumpach, Cathryn L Haigh, and Simon C Drew 18. Strain Typing of Prion Diseases using in vivo Mouse Models Aileen Boyle, Kris Hogan, Jean C. Manson, and Abigail B Diack 19. Preparation and Immunostaining of the Myenteric Plexus of Prion Infected Mice Laura J. Ellett and Victoria A. Lawson PART V: Methods for the Detection and Treatment of Prion Diseases 20. Cell Culture Methods for Screening of Prion Therapeutics Hilary E. McMahon 21. Real-Time Quaking Induced Conversion for Diagnosis of Prion Disease Katsuya Satoh, Ryuichiro Atarashi, and Noriyuki Nishida 22. Methods for Molecular Diagnosis of Human Prion Disease Jonathan D. F. Wadsworth, Gary Adamson, Susan Joiner, Lara Brock, Caroline Powell, Jacqueline M. Linehan, Jonathan A. Beck, Sebastian Brandner, Simon Mead, and John Collinge 23. Molecular sub-typing of PrPres in Human Sporadic CJD Brain Tissue Klug, G.M., Lewis, V., and Collins, S.J. Part VI: Methods for Investigation of Prion-like Disease 24. Intercellular Prion-like Conversion and Transmission of Cu/Zn Superoxide Dismutase (SOD1) in Cell Culture Leslie I. Grad, Edward Pokrishevsky, and Neil R. Cashman
