ThT 101: a primer on the use of thioflavin T to investigate amyloid formation

Kirsten Gade Malmos, Luis M. Blancas-Mejia, Benedikt Weber, Johannes Buchner, Marina Ramirez-Alvarado, Hironobu Naiki, Daniel Otzen

Research output: Contribution to journalReview articlepeer-review

73 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)1-16
Number of pages16
JournalAmyloid
Volume24
Issue number1
DOIs
StatePublished - Jan 2 2017

Keywords

  • Amyloid
  • aggregation mechanisms
  • photochemistry
  • thioflavin T

ASJC Scopus subject areas

  • Internal Medicine

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