Abstract
Seeding of amyloid fibrils into fresh solutions of the same peptide or protein in disaggregated form leads to the formation of replicate fibrils [1], with close structural similarity or identity to the original fibrillar seeds. Here we describe procedures for isolating fibrils composed mainly of β-amyloid (Aβ) from human brain and from leptomeninges, a source of cerebral blood vessels, for investigating Alzheimer’s disease and cerebral amyloid angiopathy. We also describe methods for seeding isotopically labeled, disaggregated Aβ peptide solutions for study using solid-state NMR and other techniques. These methods should be applicable to other types of amyloid fibrils, to Aβ fibrils from mice or other species, tissues other than brain, and to some non-fibrillar aggregates. These procedures allow for the examination of authentic amyloid fibrils and other protein aggregates from biological tissues without the need for labeling the tissue.
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Acknowledgments
This work was supported in part by the Intramural Research Program of the National Institute of Diabetes and Digestive and Kidney Diseases, of the National Institutes of Health and by N.I.H. grant R01 NS042852 (to S.C.M.).
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Scherpelz, K.P., Lu, JX., Tycko, R., Meredith, S.C. (2016). Preparation of Amyloid Fibrils Seeded from Brain and Meninges. In: Eliezer, D. (eds) Protein Amyloid Aggregation. Methods in Molecular Biology, vol 1345. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2978-8_20
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DOI: https://doi.org/10.1007/978-1-4939-2978-8_20
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2977-1
Online ISBN: 978-1-4939-2978-8
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