Biological Information

Background Information:

The actions of several neurotransmitters, including serotonin, are regulated through their rapid uptake and clearance from synaptic junctions by plasma membrane transport proteins. The serotonin transporter in central serotoninergic neurons is responsible for the recovery of up to 90 percent of released transmitters. The monoamine transporters are high affinity targets for a number of psychoactive agents such as cocaine, amphetamine, and antidepressants. These agents, by blocking transporters and consequently preventing neuronal uptake, elevate levels of extracellular neurotransmitter concentration in both the central and peripheral nervous systems, contributing to their behavioral and autonomic effects.

Family:

Transporter

Sub Family:

SLC6 / Monoamine

Class:

SLC

Protein Name:

SET

Uniprot Number:

P31645

Protein Aliases:

serotonin transporter 1

Gene Name:

SLC6A4

Gene ID:

6532

Gene Aliases:

5-HTT|SERT1|HTT

Accession Number:

NM_001045

Organism:

Human

Construct Details:

Full Length, Recombinant

Cell Type:

HEK293

Assay Information

Assay Type:

Functional

Assay Sub Type:

Cell Based

Functional Mode:

Antagonist

Detection Method:

Spectrofluorimetry

Measured Response:

Fluorescence

Testing Information

Procedure Summary:

Human recombinant serotonin transporter stably expressed in HEK-293 cells are used. Test compound and/or vehicle is incubated with the cells in modified Tris-HEPES buffer pH 7.1 at 25°C for 35 minutes. Alamar Blue reagent is then added to the cells in RPMI-1640 at 37°C for an additional 3 hours incubation period. Living cells will take up Alamar Blue and emit fluorescence. Fluorescence intensity is measured with excitation at 530 nm and emission at 590 nm. Decrease of 50 percent or more (≥50%) in fluorescence intensity relative to vehicle treated controls indicates significant cytotoxicity.

Incubation:

35 min at 25°C

Control Inhibitor:

Dimethyl sulfoxide

Criteria For Significance:

≥ 50% Decrease in fluorescence intensity relative to vehicle control

Test Concentration / Dose:

Compounds are screened at 10, 3, 1, 0.3, 0.1, 0.03, 0.01 and 0.003 μM

Test Sample Requirements:

75 μl of 10 mM stock or 1 mg (pre-weighed) powder (assuming FW ≦500 and purity 100%)

Minimum Order Quantity:

16

Turnaround Time

Standard:

20 Business Days

Reference Compound Data

PubChem ID

679

Name

Dimethyl sulfoxide

Measurement

23% - IC50

Clinical Relevance

Therapeutic Area:

CNS/Addiction/Pain

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei

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