Neurotransmitter transporters have become important pharmaceutical targets, as they play a key role in depression and neurodegenerative diseases (NDD) such as Alzheimers and Parkinsons diseases. The ability to monitor serotonin, norepinephrine and dopamine neurotransmitter uptake 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 this novel assay kit, researchers now have a tool to screen for live-cell 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.

 

Benefits of the Neurotransmitter Transporter Uptake Assay Kit

·               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.

 

Patented technology

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.

 

Assay Principle

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.

 

Workflow

The Neurotransmitter Transporter Uptake Assay Kit is available in Explorer and Bulk quantities, to meet the throughput needs of different users.  Please see the Order Info tab for details.

 

Performance Considerations

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.

Product Number	R8173	R8174
Product Name	Neurotransmitter Transporter Uptake Assay Explorer Kit	Neurotransmitter Transporter Uptake Assay Bulk Kit
Unit Size	1 Explorer Kit provides: 5 vials lyophilized fluorescent dye/masking dye mix Each vial provides sufficient reagent for 1 microplate (96- or 384-well)	1 Bulk Kit provides: 5 vials lyophilized fluorescent dye/masking dye mix Each vial provides sufficient reagent for 10 microplates (96- or 384-well)

To place an order, call your local Molecular Devices office or Molecular Devices distributor.