Bitan:Aggregation Other proteins (ThT Fluorescence)

α-synuclein

Protein Concentration: 100μM

Buffer: 10mM phosphate bufffer (pH 7.4)

Protocol: Dissolve α-synuclein (200 μg) in 70 μl phosphate buffer and keep at 37°C under shaking. This experiment takes ~8-10 days for completion. The ThT readings can be taken after the interval of 24h. Use the 100μl quartz cuvette for ThT measurement with α-synuclein. To 110μl of ThT (20μM in 10mM glycine-NaOH, pH 9.8) in the quartz cuvette, add 10μl of α-synuclein. Incubate for 5 min and then take ThT reading.

β2-microglobulin

Protein Concentration: 25μM

Buffer: 10mM sodium acetate (pH 3.5)

Protocol: Dissolve β2-microglobulin (137.5 μg) in 500 μl sodium acetate buffer and keep at 37°C under shaking. This experiment takes ~8-10 h for completion. The ThT readings can be taken after the interval of 1-2 h. Use the disposable cuvette for ThT measurement with β2m. To 330μl of ThT (in 10mM phosphate buffer), add 50μl of α-synuclein. Incubate for 5 min and then take ThT reading.

Calcitonin