S9C:WT (1:2) αSyn amyloid fibres were incubated with Alexa Fluor 647 maleimide dye (10‐fold molar excess, Thermo Fisher Scientific) and Biotin‐X, SE dye (2.5‐fold molar excess, Thermo Fisher Scientific) for at least 2 h at room temperature in the dark in HKMT buffer. T111C Hsc70 was incubated with Alexa Fluor 488 maleimide dye (10‐fold molar excess, Thermo Fisher Scientific) for at least 2 h at room temperature in the dark in HKMT buffer. Labelled Hsc70 was then buffer exchanged using PD SpinTrap G‐25 column (Cytiva) pre‐equilibrated in HKMD buffer. Labelled amyloid fibres (1 µM) were incubated with labelled Hsc70 (2 µM), DNAJB1 (1 µM) and Apg2 (0.2 µM, if indicated) in disaggregation buffer for 1 h at 30°C in the dark. The T111C‐Alexa Fluor 488 Hsc70 had the same disaggregation activity as unlabelled T111C‐Hsc70 (Fig EV5 ).
Alexa fluor 488 maleimide dye
Manufactured by Thermo Fisher Scientific
Alexa Fluor 488 maleimide dye is a fluorescent dye used for labeling proteins. It has an excitation maximum at 494 nm and an emission maximum at 519 nm.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using alexa fluor 488 maleimide dye
1
Labeling and Disaggregation of α-Synuclein Amyloids
2
Hsc70-mediated Amyloid Disaggregation
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S9C:WT (1:2) αSyn amyloid fibres were incubated with Alexa Fluor 647 maleimide dye (10-fold molar excess, Thermo Fisher Scientific) and Biotin-X, SE dye (2.5-fold molar excess, Thermo Fisher Scientific) for at least 2 hours at room temperature in the dark in HKMT buffer. T111C Hsc70 was incubated with Alexa Fluor 488 maleimide dye (10-fold molar excess, Thermo Fisher Scientific) for at least 2 hours at room temperature in the dark in HKMT buffer. Labelled Hsc70 was then buffer exchanged using PD SpinTrap G-25 column (Cytiva) pre-equilibrated in HKMD buffer. Labelled amyloid fibres (1 µM) were incubated with labelled Hsc70 (2 µM), DNAJB1 (1 µM) and Apg2 (0.2 µM, if indicated) in disaggregation buffer for 1 hour at 30°C in the dark.
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