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Isolering af opløselige og uopløselige PrP Oligomerer i normal human hjerne
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JoVE Journal Medicine
Isolation of Soluble and Insoluble PrP Oligomers in the Normal Human Brain

Isolering af opløselige og uopløselige PrP Oligomerer i normal human hjerne

DOI:
10.3791/3788-v

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11:29 min

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October 03, 2012

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, ,
1Department of Pathology, National Prion Disease Pathology Surveillance Center,Case Western Reserve University School of Medicine, 2Department of Neurology, National Prion Disease Pathology Surveillance Center,Case Western Reserve University School of Medicine

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

Summary

Automatic Translation

Automatic Translation

En ny art af cellulær prionprotein (PrP<sup> C</sup>) Er for nylig blevet identificeret i uinficerede humane hjerner under anvendelse af fremgangsmåderne beskrevet her. Disse fremgangsmåder kan anvendes til at isolere forskellige PrP arter, mens nogle af dem er også anvendelige til isolering af andre fejlfoldede proteinaggregater fra humane hjerner.

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

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