NET Human Norepinephrine Transporter Functional Antagonist Uptake LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
The actions of several neurotransmitters, including norepinephrine, are regulated through their rapid uptake and clearance from synaptic junctions by plasma membrane transport proteins. The norepinephrine transporter in central adrenergic neurones 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 concentrations 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:
NET
Uniprot Number:
P23975
Protein Aliases:
norepinephrine transporter
Gene Name:
SLC6A2
Gene ID:
6530
Gene Aliases:
NET|NAT1|NET1|SLC6A5
Accession Number:
XM_011523297
Organism:
Human
Construct Details:
Full Length, Recombinant
Cell Type:
MDCK
Assay Information
Assay Type:
Functional
Assay Sub Type:
Cell Based
Functional Mode:
Antagonist
Detection Method:
Radiometry
Measured Response:
Scintillation
Testing Information
Procedure Summary:
Human recombinant norepinephrine transporter expressed in MDCK cells are plated overnight. Test compound and/or vehicle is preincubated with cells in modified Tris-HEPES buffer pH 7.1 for 20 minutes at 25°C and 25 nM [³H]Norepinephrine is then added for an additional 15 minute incubation period. A lysate is obtained from solubilized cells and counted to determine [³H]Norepinephrine uptake. Reduction of [°H]Norepinephrine uptake by 50 percent or more (≥50%) relative to 10 µM desipramine indicates significant inhibitory activity. If significant inhibition of uptake is observed, item #302100-1 is suggested as follow up assay to evaluate for possible compound-induced cytotoxicity.
Incubation:
15 min at 25°C
Control Inhibitor:
Desipramine
Criteria For Significance:
≥ 50% Inhibition of [3H]Norepinephrine uptake relative to desipramine 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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