Fluorescence Methods for Unraveling Oligomeric Amyloid Intermediates

Niels Zijlstra, Nathalie Schilderink, Vinod Subramaniam

    Research output: Contribution to JournalArticleAcademicpeer-review

    Abstract

    Amyloid oligomers are considered to be the relevant toxic species in many amyloid diseases and much research effort has been devoted to fully characterize these oligomers. Despite their importance, oligomers have proven to be difficult to characterize structurally. Information on their aggregation number is scarce, largely because standard techniques struggle to provide reliable results. In this chapter, we present two different methods that reproducibly yield fluorescently labeled α-Synuclein oligomers. We then discuss a new approach, combining single-molecule photobleaching and sub-stoichiometric fluorescent labeling, that we have developed to determine the aggregation number of supramolecular protein assemblies.

    Original languageEnglish
    Pages (from-to)151-69
    Number of pages19
    JournalMethods in Molecular Biology
    Volume1345
    DOIs
    Publication statusPublished - 2016

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    Keywords

    • Fluorescent Dyes
    • Humans
    • Parkinson Disease
    • Protein Multimerization
    • Staining and Labeling
    • alpha-Synuclein
    • Journal Article
    • Research Support, Non-U.S. Gov't

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