Neurotransmitter transporters play a key role in depression and neurodegenerative diseases (NDD) such as Alzheimer's and Parkinson's. The ability to monitor serotonin, norepinephrine, and dopamine transporter (SERT, NET and DAT, respectively) activity in expressing cells, transfected or primary, is key to a better understanding of these diseases.
Until recently, radioactively labeled compounds were used to measure serotonin, norepinephrine and dopamine transporter uptake. With the introduction of the Neurotransmitter Transporter Uptake Assay Kit, researchers now have a tool to screen for live-cell kinetic uptake for these three key neurotransmitters.
- Homogeneous, live-cell fluorescent assay replaces radioactive assays, reducing disposal costs and eliminating safety risks associated with radiolabeled assays
- One-step, homogeneous assay format is amenable to automation and high-throughput screening
- Two assay setup options offer greater flexibility: endpoint mode for high-throughput applications, or real-time kinetic mode for mechanistic studies
This assay 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 is actively transported into the cells via the neurotransmitter transporters.
The Neurotransmitter Transporter Uptake Assay Kit uses a fluorescent substrate that mimics the biogenic amine neurotransmitters and is taken into the cell through those specific transporters, resulting in increased intracellular fluorescence intensity that is monitored in real time using a bottom-reading microplate reader. In this convenient no-wash procedure, cells are incubated with the kit reagent and transferred to the plate reader for evaluation. The assay is ideal for use in academic, biotech, and pharmaceutical environments for research, assay development, and screening.
For more details, please refer to the Neurotransmitter Transporter Uptake Assay Kit product insert, which may be accessed through the Molecular Devices Knowledge Base.
Solvent Tolerance: This assay can tolerate a final solvent concentration of up to 1% DMSO or Ethanol with minimal impact on the precision or the overall assay performance in the cell lines tested.
Cell Density: Density of the cells and cell plating could influence the assay performance as well as the signal readout. Therefore, it is recommended that the cell plating information supplied in the product insert be used as a guideline and that further optimization be performed to maximize assay performance.
Same-day Cell Plating: Cells can be plated on the same day that the assay is run without compromising the assay performance or precision. However, it is advised that the cell number and the amount of time for the cells to adhere to the plate be optimized for the specific cell line.
BSA Addition to HBSS Buffer: Similar performance is seen with or without the addition of BSA. However, the use of BSA can help reduce binding of compounds to the walls of the plate well or pipet tips.
Order of Reagent Additions: Changing the order of reagent additions, i.e., dye before inhibitor, may result in a reduced signal. Therefore, following the suggested protocol and guidelines is recommended to ensure the desired assay performance.
Nisoxetine inhibition in HEK-hSERT cells
IC50 correlation for DAT, NET and SERT (customer results)
Neurotransmitter Transporter Uptake Assay Kit is compatible with these systems:
- FLIPR Tetra High Throughput Cellular Screening System
- FlexStation 3 Benchtop Multi-Mode Microplate Reader
- SpectraMax i3 Multi-Mode Detection Platform
- SpectraMax M5e/M5/M4/M3/M2e Microplate Readers
- SpectraMax Paradigm Multi-Mode Microplate Reader
- Gemini EM Microplate Reader
For more information, view the assay kits & instrument compatibility table.
|Product Name||Description||Part Number|
Neurotransmitter Transporter Uptake Assay Kit
1 vial = 1 plate x 2 vials each
1 vial = 1 plate x 5 vials
1 vial = 10 plates x 5 vials
Compatible with 96- and 384-well plate formats.