title: ThT 101: a primer on the use of thioflavin T to investigate amyloid formation.
abstract: Thioflavin T (ThT) has been widely used to investigate amyloid formation since 1989. While concerns have recently been raised about its use as a probe specific for amyloid, ThT still continues to be a very valuable tool for studying kinetic aspects of fibrillation and associated inhibition mechanisms. This review aims to provide a conceptual instruction manual, covering appropriate considerations and pitfalls related to the use of ThT. We start by giving a brief introduction to amyloid formation with focus on the morphology of different aggregate species, followed by a discussion of the quality of protein needed to obtain reliable fibrillation data. After an overview of the photochemical basis for ThT's amyloid binding properties and artifacts that may arise from this, we describe how to plan and analyze ThT assays. We conclude with recommendations for complementary techniques to address shortcomings in the ThT assay.
authors.affiliation: a Interdisciplinary Nanoscience Center (iNANO) and Center for Insoluble Protein Structures (inSPIN) , Aarhus University , Aarhus C , Denmark.|b Department of Molecular Biology and Genetics , Aarhus University , Aarhus C , Denmark.
authors.forename: Kirsten
authors.initials: K
authors.lastname: Gade Malmos
authors.affiliation: c Department of Biochemistry and Molecular Biology , Mayo Clinic , Rochester , MN , USA.
authors.forename: Luis M
authors.initials: LM
authors.lastname: Blancas-Mejia
authors.affiliation: d Center for Integrated Protein Science Munich at the Department Chemie , Technische Universität München , Garching , Germany.
authors.forename: Benedikt
authors.initials: B
authors.lastname: Weber
authors.affiliation: d Center for Integrated Protein Science Munich at the Department Chemie , Technische Universität München , Garching , Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Buchner
authors.affiliation: c Department of Biochemistry and Molecular Biology , Mayo Clinic , Rochester , MN , USA.
authors.forename: Marina
authors.initials: M
authors.lastname: Ramirez-Alvarado
authors.affiliation: e Department of Molecular Pathology, Faculty of Medical Sciences , University of Fukui , Fukui , Japan.
authors.forename: Hironobu
authors.initials: H
authors.lastname: Naiki
authors.affiliation: a Interdisciplinary Nanoscience Center (iNANO) and Center for Insoluble Protein Structures (inSPIN) , Aarhus University , Aarhus C , Denmark.
authors.forename: Daniel
authors.identifier: http://orcid.org/0000-0002-2918-8989
authors.initials: D
authors.lastname: Otzen
mesh_terms: D000682:Amyloid / Q000737:chemistry*; D000818:Animals; D052160:Benzothiazoles; D006801:Humans; D010777:Photochemistry / Q000379:methods*; D013844:Thiazoles / Q000737:chemistry*
chemical_list: D000682:Amyloid; D052160:Benzothiazoles; D013844:Thiazoles; C009462:thioflavin T
keywords: Amyloid; aggregation mechanisms; photochemistry; thioflavin T