SERT Human Serotonin Transporter Functional Antagonist Uptake LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
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:
Radiometry
Measured Response:
Scintillation
Testing Information
Procedure Summary:
HEK-293 cells expressing human recombinant serotonin transporter are plated. Test compound and/or vehicle is preincubated with cells (2 x 10⁵/ml) in modified Tris-HEPES buffer pH 7.1 for 20 minutes at 25°C and 65 nM [3H]Serotonin is then added for an additional 15-minute incubation period. Bound cells are filtered and counted to determine [3H]Serotonin uptake. Reduction of [3H]Serotonin uptake by 50 percent or more (≥50%) relative to 1 μM fluoxetine indicates significant inhibitory activity. If significant inhibition of uptake is observed, item #364100-1 is suggested as a follow-up assay to evaluate for possible compound-induced cytotoxicity.
Incubation:
15 min at 25°C
Control Inhibitor:
Fluoxetine
Criteria For Significance:
≥ 50% Inhibition of [3H]Serotonin uptake relative to fluoxetine response
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
Clinical Relevance
Therapeutic Area:
CNS/Addiction/Pain
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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