nAChR (alpha3/beta4) Human Ion Channel Binding Agonist Radioligand LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Nicotinic acetylcholine receptors (nAChRs) are a diverse family of neurotransmitter-gated ion channels, which contain five transmembrane subunits arranged around a central pore. Their amino acid sequences lie in the same superfamily as the GABA, glycine, serotonin 5-HT3 and ionic glutamate receptors, within the general class of ligand-gated ion channels. Selected nAChRs mediate classical excitatory neurotransmission at the neuromuscular junction, autonomic ganglia and selected synapses in the brain and spinal cord. Receptors are also expressed in a variety of tissue and cell types: lymphocytes, fibroblasts, pulmonary neuroendocrine cells, spermatozoa, granulocytes, etc. Nicotinic acetylcholine receptor agonism may improve learning and memory, increase dopaminergic transmission and exhibit analgesic activity. Receptor agonism may also stimulate autonomic cardiovascular, gastrointestinal function including nausea and emesis, salivary and bronchial secretion and facilitate convulsions.
Family:
Ion Channel
Sub Family:
Acetylcholine (Nicotinic)
Class:
Ligand-Gated
Protein Name:
nAChR-a3b4
Uniprot Number:
P32297, P30926
Gene Name:
CHRNA3, CHRNB4
Gene ID:
1136, 1143
Organism:
Human
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Binding
Detection Method:
Radiometry
Testing Information
Procedure Summary:
Human recombinant nicotinic acetylcholine alpha3/beta4 receptors expressed in CHO-K1 cells are used in modified Tris-HCl buffer pH 7.4. A 0.6 µgǂ aliquot incubated with 0.05 nM [¹²⁵I]Epibatidine for 60 minutes at 25℃. Non-specific binding is estimated in the presence of 10 µM Epibatidine. Membranes are filtered and washed, the filters are then counted to determine [¹²⁵I]Epibatidine specifically bound. Compounds are screened at 10 µM.
Note: ǂMembrane may change from lot to lot, the concentration used will be adjusted if necessary.
Ligand:
[¹²⁵I] Epibatidine
Ligand Kd (nM):
0.13
Ligand Concentration:
0.05 nM
Non Specific:
10 µM Epibatidine
Incubation:
60 min at 25°C
Control Inhibitor:
(-)-Epibatidine
Criteria For Significance:
≥ 50% of max stimulation or inhibition
Test Sample Requirements:
Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).
Minimum Order Quantity:
2
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Clinical Relevance
Therapeutic Area:
CNS/Addiction/Pain
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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