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A filter retardation assay facilitates the detection and quantification of heat-stable, amyloidogenic mutant huntingtin aggregates in complex biosamples

Item Type:Article
Title:A filter retardation assay facilitates the detection and quantification of heat-stable, amyloidogenic mutant huntingtin aggregates in complex biosamples
Creators Name:Ast, A. and Schindler, F. and Buntru, A. and Schnoegl, S. and Wanker, E.E.
Abstract:N-terminal mutant huntingtin (mHTT) fragments with pathogenic polyglutamine (polyQ) tracts spontaneously form stable, amyloidogenic protein aggregates with a fibrillar morphology. Such structures are detectable in brains of Huntington's disease (HD) patients and various model organisms, suggesting that they play a critical role in pathogenesis. Heat-stable, fibrillar mHTT aggregates can be detected and quantified in cells and tissues using a denaturing filter retardation assay (FRA). Here, we describe step-by-step protocols and experimental procedures for the investigation of mHTT aggregates in complex biosamples using FRAs. The methods are illustrated with examples from studies in cellular, transgenic fly, and mouse models of HD, but can be adapted for any disease-relevant protein with amyloidogenic polyQ tracts.
Keywords:Filter Retardation Assay, Protein Misfolding, Protein Aggregation, Coaggregation, Insoluble Aggregates, Amyloidogenesis, Seeding and Spreading of Protein Aggregates, Animals, Mice
Source:Methods in Molecular Biology
Series Name:Methods in Molecular Biology
Title of Book:Huntington’s Disease
ISSN:1064-3745
ISBN:978-1-4939-7825-0
Publisher:Springer / Humana Press (U.S.A.)
Volume:1780
Page Range:31-40
Date:2018
Official Publication:https://doi.org/10.1007/978-1-4939-7825-0_3
PubMed:View item in PubMed

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