How does the thioflavin T assay work?
How does the thioflavin T assay work?
Thioflavin T (ThT) is a commonly used probe to monitor in vitro amyloid fibril formation. Upon binding to amyloid fibrils, ThT gives a strong fluorescence signal at approximately 482 nm when excited at 450 nm [2].
What is Thioflavin T used for?
Thioflavin T (ThT) dye fluorescence is used regularly to quantify the formation and inhibition of amyloid fibrils in the presence of anti-amyloidogenic compounds such as polyphenols.
How does Thioflavin T bind to amyloid fibrils?
This suggests that positive charge on the thioflavin T molecule has a role in its micelle formation that then bind the amyloid fibrils. Our data suggests that the micelles of thioflavin T bind amyloid fibrils leading to enhancement of fluorescence emission.
Is Thioflavin T light sensitive?
In contrast, the fluorochrome dyes, thioflavin S and T, exhibit a dramatic increase in fluorescent brightness upon binding to amyloid. The binding of thioflavin dyes is linked to the presence of cross-β structure in the amyloid fibrils, and this stain is very sensitive.
What is the difference between Thioflavin T and S?
Thioflavin S Like thioflavin T it binds to amyloid fibrils but not monomers and gives a distinct increase in fluorescence emission. However unlike thioflavin T, it does not produce a characteristic shift in the excitation or emission spectra.
How do you make Thioflavin T solution?
Preparation of Thioflavin T (ThT): Prepare 10 mM ThT stock (Dissolve 31.8 mg of ThT in 10 ml of Tris buffer, pH 8.0). Add 5% sodium azide (NaN3) to this solution so as the final NaN3 concentration is 0.01%. The solution is freshly prepared and filtered through 0.2 micron sterile syringe filter before use.
How do you dissolve Thioflavin T?
Is Thioflavin toxic?
Thioflavins are corrosive, irritants, and are acutely toxic, causing serious eye damage. Thioflavin T has been used in research into Alzheimer’s disease and other neurodegenerative diseases.
What is the color of Thioflavin T?
Basic Yellow 1
Thioflavin T (Basic Yellow 1, Methylene yellow, CI 49005, or ThT) is a benzothiazole salt obtained by the methylation of dehydrothiotoluidine with methanol in the presence of hydrochloric acid.
What are amyloid fibrils made of?
Amyloid fibrils are formed by normally soluble proteins, which assemble to form insoluble fibers that are resistant to degradation. Their formation can accompany disease and each disease is characterized by a specfic protein or peptide that aggregates.
