Nonspecific binding was determined by carrying out parallel determinations in the presence of excess unlabeled competitive antagonist (10 µM naloxone).
Non-specific binding was determined by co-incubation with excess Nisoxetine (10 μM).
Non-specific binding was determined by adding 100-nM solution of unlabeled GLP-1.
Plasma protein binding was determined by using human plasma according to Patila et al. (2007).
Total and nonspecific radioligand binding was determined by reaction without and with 10 μM thioperamide and clobenpropit, respectively.
The non-displaceable binding was determined by addition of 10 to 20 μM tetrabenazine (BIOTREND Chemikalien GmbH, Cologne, Germany).
The level of specific binding was determined by comparing brain uptake in the KOR-enriched striatum region, representing total binding, with the KOR-free cerebellum region, representing NSB.
The effect of these mutations on membrane binding was determined by a quantitative phospholipid ELISA assay and compared to wild-type α-Syn and to the Parkinson's disease-causing mutations, A30P, E46K and A53T.
Specific binding was determined by subtracting values of non-specific binding from that of total binding.
Non-specific binding was determined by measuring bound radioactivity in the absence of Ir-LBP.
Nonspecific binding was determined by using 1 µM cold CCK-8 under the same incubation conditions above.
