Acetylcholinesterase (AChE) activity was measured following a procedure described previously [13 (link)]. Briefly, EV samples (100 µL) were mixed with 1.25mM acetylthiocholine in PBS at pH 8 plus 0.1mM 5,5-dithio-bis (2-nitrobenzoic acid) in PBS at pH 7. The resulting suspension (200 µL) was held at room temperature and then warmed to 37°C for 10 minutes for optical density reading. Absorption at 450 nm was monitored for 10 minutes using a plate reader spectrophotometer (Molecular Devices with SOFTmax® Pro version 5, San Jose, CA, USA).
Bazié W.W., Boucher J., Vitry J., Goyer B., Routy J.P., Tremblay C., Trottier S., Jenabian M.A., Provost P., Alary M, & Gilbert C. (2021). Plasma Extracellular Vesicle Subtypes May be Useful as Potential Biomarkers of Immune Activation in People With HIV. Pathogens and Immunity, 6(1), 1-28.
Other organizations :
Centre Muraz, Centre hospitalier de l'Université Laval, Université Laval, McGill University Health Centre, Université de Montréal, Centre Hospitalier de l’Université de Montréal, Université du Québec à Montréal, Institut National de Santé Publique du Québec
Concentration of 5,5-dithio-bis (2-nitrobenzoic acid) (0.1mM)
PH of 5,5-dithio-bis (2-nitrobenzoic acid) solution (pH 7)
Temperature of the reaction (37°C)
Duration of the reaction (10 minutes)
Spectrophotometer used (Molecular Devices with SOFTmax® Pro version 5)
controls
None explicitly mentioned
Annotations
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