Quantification of Oxidized Dopamine

The near-infrared fluorescence assay was performed as described previously (6 (link), 27 (link)). Leftover pellets from SDS extraction were extracted in 1 M NaOH at 55°C overnight. Then, samples were dried, washed with H₂O, lyophilized, and solubilized in H₂O. Samples and standard solutions from a 10 mM oxidized dopamine stock were dropped on a Biodyne Nylon Transfer Membrane (Pall). Membranes were scanned using Odyssey infrared imaging system (LI-COR) with the 700 channel. Samples were quantified by obtaining integrated spot intensities using Odyssey infrared imaging software (LI-COR).

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Burbulla L.F., Zheng J., Song P., Jiang W., Johnson M.E., Brundin P, & Krainc D. (2021). Direct targeting of wild-type glucocerebrosidase by antipsychotic quetiapine improves pathogenic phenotypes in Parkinson’s disease models. JCI Insight, 6(19), e148649.

Publication 2021

Odyssey infrared imaging system Odyssey infrared imaging software

Assay Dopamine Fluorescence Membranes Nylon Pellets

Corresponding Organization : Northwestern University

Other organizations : German Center for Neurodegenerative Diseases, Munich Cluster for Systems Neurology, Ludwig-Maximilians-Universität München, Van Andel Institute

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Variable analysis

independent variables

Extraction in 1 M NaOH at 55°C overnight

dependent variables

Fluorescence intensity of samples

control variables

Biodyne Nylon Transfer Membrane (Pall)
Odyssey infrared imaging system (LI-COR) with the 700 channel
Odyssey infrared imaging software (LI-COR)

controls

Standard solutions from a 10 mM oxidized dopamine stock

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