Biological Origin: Human. Biological Activity: 100 µM A53T alpha synuclein protein monomer seeded with 10 uM A53T alpha synuclein protein PFF in 25 µM Thioflavin T (PBS pH 7.4, 100 µl reaction volume) generated a fluorescence intensity of 28 000 Relative Fluorescence Units after incubation at 37C with shaking at 600 rpm for 56 hours. Fluorescence was measured by excitation at 450 nm and emission at 485 nm on a Molecular Devices Gemini XPS microplate reader. Application Notes: Certified >95% pure using SDS-PAGE analysis. Low endotoxin <5 EU/mL 2mg/mL
SDS-PAGE of ~14 kDa A53T Alpha Synuclein Monomer
Thioflavin T is a fluorescent dye that binds to beta sheet-rich structures such as those in alpha synuclein fibrils. Upon binding, the emission spectrum of the dye experiences a red-shift and increased fluorescence intensity. Thioflavin T emission curves show a limited increase in fluorescence (correlated to alpha synuclein aggregation) over time in A53T alpha synuclein monomers. A much greater increase in fluorescence is seen when 100 uM monomer is combined with 10 nM of fibrils as the fibrils seed the formation of new fibrils from the pool of active monomers. Thioflavin T ex = 450 nm, em = 485 nm.
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