Assay Method Information

Assay Name:  Receptor Binding Assay
Description:  Receptor binding assays for compounds determined by LC-MS to be >95% pure were performed by the Psychoactive Drug Screening Program (PDSP) at Chapel Hill, North Carolina (Besnard et al, 2012). Briefly, binding affinities, Ki, for σ2R/TMEM97 (rat PC12 cells) were determined through competition binding assays using the radioligand [3H]-ditolylguanidine in the presence of (+)-pentazocine to block σ1R binding sites, whereas binding affinities, Ki, for σ1R (guinea pig brain) were determined through competition binding assays with [3H]-(+)-pentazocine. Binding affinities, Ki, for σ2R/TMEM97 (human clone transiently expressed in HEK293 cells) were determined through competition binding assays using the radioligand [3H]-ditolylguanidine in the presence of (+)-pentazocine to block σ1R binding sites, and binding affinities, Ki, for σ1R (human clone transiently expressed in HEK293 cells) were determined through competition binding assays with [3H]-(+)-pentazocine. Ki values are calculated from best-fit IC50 determinations performed in triplicate. For those σ2R/TMEM97 modulators that were found to be neuroprotective in these studies, their Ki values for other CNS proteins were also determined by the PDSP.
Affinity data for this assay
 

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