ThT 101: a primer on the use of thioflavin T to investigate amyloid formation, 2017, Malmos et al.

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

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.

Link | PDF (Amyloid)

 

[109] is Dye-Binding Assays for Evaluation of the Effects of Small Molecule Inhibitors on Amyloid Aβ Self-Assembly (2012, ACS Publications)

Quercetin, resveratrol and EGCG are common supplements taken by patients so there's a possibility that might reduce the apparent amyloid signal.

 

Interesting. I wonder if small molecules may be more of a problem in fluid phase assays. For tissue staining a lot of small molecules may be washed out of the tissue by washing or incubation steps that can be quite lengthy.