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Prion Protein Protocols brings together a collection of current protocols in the field of mammalian prion disease research. Since identification of the prion protein gene some 20 years ago, what were once thought to be rare, neurodegenerative diseases of humans and animals have become a major research area. The major interest in this field results from the epidemic of bovine spongiform encephalopathy and the subsequent human prion disease, variant Creutzfeldt-Jakob disease, which still poses an unknown risk to human health. The "unknowns" in the field stem from the enigmatic nature of infectious prions, the infectious agent that can transmit these diseases between individuals. Although much information has been gained over the past two decades about the molecular nature of prion proteins, only very recently have several research groups begun to cast light on how to turn the normal prion protein into its aberrant, infectious form. Prion Protein Protocols brings together a collection of protocols from 13 diff- ent laboratories in five countries covering basic science and diagnostic areas of prion research. Together, the chapters provide an up-to-date collection of current methods in this unique area of neuroscience. The notes section at the end of each methods chapter provides useful insight into the experimental techniques, and they are no doubt a benefit to researchers wanting to use these technologies.
List of contents
Cell Culture Models to Unravel Prion Protein Function and Aberrancies in Prion Diseases.- Investigation of PrPC Metabolism and Function in Live Cells.- Immunodetection of PrPSc Using Western and Slot Blotting Techniques.- Assaying Prions in Cell Culture.- Generation of Cell Lines Propagating Infectious Prions and the Isolation and Characterization of Cell-derived Exosomes.- Neurotoxicity of Prion Peptides on Cultured Cerebellar Neurons.- Understanding the Nature of Prion Diseases Using Cell-free Assays.- Methods for Conversion of Prion Protein into Amyloid Fibrils.- Amplification of Purified Prions In Vitro.- Expression and Purification of Full-Length Recombinant PrP of High Purity.- Analysis of PrP Conformation Using Circular Dichroism.- Effect of Copper on the De Novo Generation of Prion Protein Expressed in Pichia pastoris.- Biophysical Investigations of the Prion Protein Using Electron Paramagnetic Resonance.- Molecular Diagnosis of Human Prion Disease.- Analysis of Endogenous PrPC Processing in Neuronal and Non-neuronal Cell Lines.- Molecular Typing of PrPres in Human Sporadic CJD Brain Tissue.- Transgenic Mouse Models of Prion Diseases.- Quantitative Bioassay of Surface-bound Prion Infectivity.
Summary
Prion Protein Protocols brings together a collection of current protocols in the field of mammalian prion disease research. Since identification of the prion protein gene some 20 years ago, what were once thought to be rare, neurodegenerative diseases of humans and animals have become a major research area. The major interest in this field results from the epidemic of bovine spongiform encephalopathy and the subsequent human prion disease, variant Creutzfeldt–Jakob disease, which still poses an unknown risk to human health. The “unknowns” in the field stem from the enigmatic nature of infectious prions, the infectious agent that can transmit these diseases between individuals. Although much information has been gained over the past two decades about the molecular nature of prion proteins, only very recently have several research groups begun to cast light on how to turn the normal prion protein into its aberrant, infectious form. Prion Protein Protocols brings together a collection of protocols from 13 diff- ent laboratories in five countries covering basic science and diagnostic areas of prion research. Together, the chapters provide an up-to-date collection of current methods in this unique area of neuroscience. The notes section at the end of each methods chapter provides useful insight into the experimental techniques, and they are no doubt a benefit to researchers wanting to use these technologies.
Additional text
From the reviews:
"Interspersed are concise reviews focusing on current growth areas in the prion field. … The protocols are clear and authoritative. For those entering the field it would be an excellent guide." (Hugh Pennington, Microbiology Today, February, 2009)
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From the reviews:
"Interspersed are concise reviews focusing on current growth areas in the prion field. ... The protocols are clear and authoritative. For those entering the field it would be an excellent guide." (Hugh Pennington, Microbiology Today, February, 2009)
