Optotracing using Amytracker

Amytracker are optotracers with structure-dependent photo-physical properties. All Amytracker variants are designed to bind to the Congo red binding pocket on the amyloid fibril and require a theoretical minimum of eight in-register parallel-β-strands for binding. Therefore, Amytracker reliably labels amyloids derived from a variety of amyloidogenic proteins or peptides from different species. Due to their structure-dependent photo-physical properties, the Amytracker variants are only fluorescent when binding to a target and different targets can produce a difference in the molecules fluorescence spectrum. To investigate different targets, we recommend collecting excitation and emission spectra with excitation and emission parameters summarised in the table below. Reference spectra for all Amytracker variants can be accessed here.

Table: Amytracker spectral properties with maximum excitation (Exmax) and emission (Emmax) when bound and recommended range for acquisition of excitation- and emission spectra as well as recommended filter sets for microscopy.
Exmax Emmax Excitation spectrum (detect at Emmax) Emission spectrum (excite at Exmax) Recommended
Amytracker 480 420 nm 480 nm 300 - 450 nm 450 - 800 nm DAPI
Amytracker 520 460 nm 520 nm 300 - 490 nm 490 - 800 nm FITC, GFP
Amytracker 540 480 nm 540 nm 300 - 510 nm 510 - 800 nm FITC, GFP, YFP
Amytracker 630 520 nm 630 nm 300 - 600 nm 550 - 800 nm PI, Cy3, TxRed, mCherry, Cy3.5
Amytracker 680 530 nm 680 nm 300 - 650 nm 660 - 800 nm PI, mCherry, Cy3.5

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