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 PrP^Sc 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 Andréoletti, 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. Orrù, 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 Fernández-Borges, Hasier Eraña, Saioa Rodríguez-Elezgarai, Chafik Harrathi, Vanesa Venegas, and Joaquín 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 PrP^res 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