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Label-free high-throughput screening assay for the identification of norepinephrine transporter (NET/SLC6A2) inhibitors | Scientific Reports
Description
The human norepinephrine transporter (NET) is an established drug target for a wide range of psychiatric disorders. Conventional methods that are used to functionally characterize NET inhibitors are based on the use of radiolabeled or fluorescent substrates. These methods are highly informative, but pose limitations to either high-throughput screening (HTS) adaptation or physiologically accurate representation of the endogenous uptake events. Recently, we developed a label-free functional assay based on the activation of G protein-coupled receptors by a transported substrate, termed the TRACT assay. In this study, the TRACT assay technology was applied to NET expressed in a doxycycline-inducible HEK 293 JumpIn cell line. Three endogenous substrates of NET—norepinephrine (NE), dopamine (DA) and epinephrine (EP)—were compared in the characterization of the reference NET inhibitor nisoxetine. The resulting assay, using NE as a substrate, was validated in a manual HTS set-up with a Z′ = 0.55. The inhibitory potencies of several reported NET inhibitors from the TRACT assay showed positive correlation with those from an established fluorescent substrate uptake assay. These findings demonstrate the suitability of the TRACT assay for HTS characterization and screening of NET inhibitors and provide a basis for investigation of other solute carrier transporters with label-free biosensors.#Sport & Camping
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# Sport & Camping
Updated
10 June 2021
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Increase your cardio and keep fit at home with these affordable treadmills - Technology News
Description
Fitness is important, however, we often do not get time due to our busy work schedule to visit a gym or go for outdoor activities. Having a treadmill at home can help you save time and complete your cardio goals. In this article, we list the best affordable treadmills for workouts.#Sport & Camping
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# Sport & Camping
Updated
07 June 2021