TY - JOUR
T1 - ThT 101
T2 - a primer on the use of thioflavin T to investigate amyloid formation
AU - Malmos, Kirsten
AU - Blancas-Mejia, Luis M
AU - Weber, Benedikt
AU - Buchner, Johannes
AU - Ramirez-Alvarado, Marina
AU - Naiki, Hironobu
AU - Otzen, Daniel
PY - 2017/1/2
Y1 - 2017/1/2
N2 - 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.
AB - 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.
KW - Journal Article
UR - http://www.scopus.com/inward/record.url?scp=85017265343&partnerID=8YFLogxK
U2 - 10.1080/13506129.2017.1304905
DO - 10.1080/13506129.2017.1304905
M3 - Journal article
C2 - 28393556
SN - 1350-6129
VL - 24
SP - 1
EP - 16
JO - Amyloid: the Journal of Protein Folding Disorders
JF - Amyloid: the Journal of Protein Folding Disorders
IS - 1
ER -