Neurotransmitter Transporter Uptake Assay Kit A live-cell kinetic assay to measure uptake of serotonin, norepinephrine, and dopamine neurotransmitters KEY FEATURES • Homogeneous, live-cell fluorescent assay • Convenient, single reagent addition assay format • No use of radioactivity required • Amenable to automation and HTS • Analyze on any bottom-read fluorescent microplate instrument • Read in endpoint mode for high-throughput applications • Read in real-time kinetic mode for mechanistic studies Introduction Neurotransmitter transporters have become important pharmaceutical targets, as they play a key role in depression and neurodegenerative diseases (NDD) such as Alzheimer’s and Parkinson’s diseases. The ability to monitor serotonin, norepinephrine and dopamine neurotransmitter uptake is key to a better understanding of these diseases. Until now, radioactively labeled compounds were used to measure serotonin, norepinephrine and dopamine transporter uptake. With the introduction of this novel assay kit, researchers will now have a tool to screen for livecell kinetic uptake for these three key neurotransmitters in a homogeneous, fluorescence-based, high-throughput format. The assay can be performed either as a real-time kinetic assay suitable for mechanistic studies or as an HTS-amenable endpoint read. Homogeneous, simple, and flexible The homogeneous Neurotransmitter Transporter Uptake Assay has a flexible and simple mix-and-read protocol that can be performed in 96- or 384-well microplates for detection with either kinetic or endpoint modes. It is ideal for use in research, assay development and high-throughput screening applications. This assay improves on current transport and binding assays for these three neurotransmitter transporters, which use radiolabeled ligands, require expensive, specialized equipment, and only detect endpoints. With this assay, researchers now have the ability to measure live-cell kinetics of these important transporters without reliance upon radioactivity and without being limited to endpoint read-only assays.

neurotransmitter iitating fluorophoi “urotransmitter transporter neurotransmitter n itating fluorophore neurotransmitt transporter Patented technology This assay, like all Molecular Devices cell-based assays, utilizes a Molecular Devices masking dye to extinguish extracellular fluorescence. This masking dye technology has been exclusively licensed from Bayer AG (U.S. Patent Nos. 6,420,183, 7,063,952, 7,138,280 and European Patent No. 0,906,572). The assay also includes a novel proprietary fluorescent indicator dye that mimics the neurotransmitters serotonin, norepinephrine, and dopamine, and...

FLIPR® Tetra System Fluorescent transporter assay (log app K,) Figure 4. Correlation of IC50 values for fluoresent assay vs. published values. Scatter plot correlating apparent Kj determined with the neurotransmitter transporter assay with literature values for known inhibitors of dopamine (DAT), serotonin (SERT), and norepinephrine (NET) re-uptake. The combined correlation coefficient R2 is greater than 0.9 indicating good correlation between literature and experimental values for this kit. Assay performance considerations Solvent tolerance This assay can tolerate a final solvent concentration...