title: Detection and Quantification Methods for Fibrillar Products of In Vitro Tau Aggregation Assays.
abstract: Alzheimer's disease is characterized in part by the intracellular misfolding and aggregation of tau protein. The aggregates, which range in size from small oligomers to large filaments, are markers for disease diagnosis and staging, potential vectors for disease propagation, and candidate sources of neurotoxicity. Here we present protocols for synthesizing large tau aggregates characterized by filamentous morphology and cross-β-sheet structure from monomeric full-length tau precursors in vitro. We also describe their detection and quantification through thioflavin dye binding, filter trap, and transmission electron microscopy methods. These methods cover applications requiring high-throughput capability as well as those requiring high-resolution analysis of aggregation mechanism.
authors.affiliation: Neuroscience Signature Program, The Ohio State University, Columbus, OH, USA.
authors.forename: Niki
authors.initials: N
authors.lastname: Nanavaty
authors.affiliation: Neuroscience Signature Program, The Ohio State University, Columbus, OH, USA.
authors.forename: Lauren
authors.initials: L
authors.lastname: Lin
authors.affiliation: Ohio State Biochemistry Program, The Ohio State University, Columbus, OH, USA.
authors.forename: Samantha H
authors.initials: SH
authors.lastname: Hinckley
authors.affiliation: Ohio State Biochemistry Program, The Ohio State University, Columbus, OH, USA. kuret.3@osu.edu.|Department of Biological Chemistry and Pharmacology, The Ohio State University College of Medicine, 1060 Carmack Rd., Columbus, OH, 43210, USA. kuret.3@osu.edu.
authors.forename: Jeff
authors.initials: J
authors.lastname: Kuret
mesh_terms: D000544:Alzheimer Disease / Q000378:metabolism; D000818:Animals; D052160:Benzothiazoles; D001681:Biological Assay / Q000379:methods*; D006801:Humans; D008854:Microscopy, Electron; D066263:Protein Aggregation, Pathological / Q000378:metabolism*; D013844:Thiazoles / Q000737:chemistry; D016875:tau Proteins / Q000032:analysis* / Q000737:chemistry* / Q000648:ultrastructure
chemical_list: D052160:Benzothiazoles; D013844:Thiazoles; D016875:tau Proteins; C009462:thioflavin T
keywords: Aggregation; Alzheimer’s disease; Electron microscopy; Filter trap assay; Tau; Thioflavin dye saturation assay