Sedimentation Velocity Analysis of Amyloid Fibrils
Chi L.L. Pham, Yee-Foong Mok, Geoffrey J. Howlett
Abstract
Chi L.L. Pham, Yee-Foong Mok, Geoffrey J. Howlett
Abstract
Analytical ultracentrifugation is a classical technique used to study the solution behavior of proteins. Experimentally determined sedimentation coefficients provide information regarding the size, shape, and interactions of biological macromolecules. Sedimentation velocity methods have been used to characterize the different aggregation states of amyloid oligomers and fibrils. This chapter first describes the theoretical background for sedimentation velocity analysis. It then provides experimental protocols for sedimentation velocity experiments using the analytical ultracentrifuge. Finally, this chapter describes the procedure used to analyze sedimentation velocity data to obtain the size distribution of amyloid fibrils and their oligomeric precursors.
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Analytical ultracentrifugation is a classical technique used to study the solution behavior of proteins. Experimentally determined sedimentation coefficients provide information regarding the size, shape, and interactions of biological macromolecules. Sedimentation velocity methods have been used to characterize the different aggregation states of amyloid oligomers and fibrils. This chapter first describes the theoretical background for sedimentation velocity analysis. It then provides experimental protocols for sedimentation velocity experiments using the analytical ultracentrifuge. Finally, this chapter describes the procedure used to analyze sedimentation velocity data to obtain the size distribution of amyloid fibrils and their oligomeric precursors.
Key concepts: Analytical Ultracentrifugation, Sedimentation, Ultracentrifuge, Sedimentation coefficient, Fibril, Amyloid fibril, Amyloid (mycology), Sedimentation equilibrium