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Chapters
- 00:05Title
- 02:02Preparation of Brain Homogenate and Detergent Soluble and Insoluble Fractions
- 03:21Velocity Sedimentation in Sucrose Step Gradients
- 04:30Size Exclusion Chromatography
- 06:16Capture of PrP with g5P and Western Blotting
- 08:20Representative PrP Isolation Results
- 10:45Conclusion
A new species of cellular prion protein (PrPC) has recently been identified in uninfected human brains using the methods described here. These methods can be used to isolate various PrP species, while some of them are also useful in isolating other misfolded protein aggregates from human brains.
Tags
Soluble PrP OligomersInsoluble PrP OligomersNormal Human BrainPrPCPrPScDetergent-solubleProteinase K (PK)-digestionDetergent-insoluble AggregatesConformational Transitionα-helical Structuresβ-sheet StructuresEndogenous PrPScIntermediate PrPSilent PrionBiophysical ApproachesBiochemical ApproachesUltracentrifugationSucrose Step Gradient SedimentationSize Exclusion ChromatographyIPrP EnrichmentGene 5 Protein (g5p)PK-treatment
