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Pharmacology JoVE (Open Access) Citable · DOI

In Vitro Assay for Studying the Aggregation of Tau Protein and Drug Screening

DOI: 10.3791/58570-v
What you'll learn
  • ✓ Set up and run a ThT-based tau aggregation assay using a microplate reader
  • ✓ Interpret fluorescence kinetics to characterize tau protein misfolding
  • ✓ Apply in vitro aggregation data for drug screening and candidate evaluation
Protocol

The tau aggregation assay described in this manuscript mimics the anticipated features of in vivo tau misfolding and aggregation.

Difficulty
intermediate
Total time
~4–6 hours per assay run (including incubation time)
Biosafety
BSL-1

Steps

1
Prepare continuous-mode ThT aggregation assay

Set up tau protein samples with thioflavin T (ThT) fluorescent dye in a multi-mode microplate reader. Configure continuous kinetic monitoring to track real-time tau aggregation via fluorescence emission.

▶ 00:45
2
Analyze tau aggregation kinetics and results

Examine fluorescence intensity curves to quantify tau protein aggregation kinetics. Compare control and drug-treated samples to assess inhibition or promotion of misfolding.

▶ 06:43
3
Interpret findings for drug screening applications

Summarize how in vitro tau aggregation assay data can predict in vivo tau misfolding behavior and support candidate drug identification.

▶ 08:50
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