Biological Information
Background Information:
Serotonin 5-HT receptors belong to the superfamily of G protein-coupled seven transmembrane proteins with the exception of the 5-HT3 subtype, which belongs to the ligand gated cation superfamily of receptors. G-protein-coupled receptors constitute one of the major signal transduction systems in eukaryotic cells. Coding sequences for these receptors, in those regions believed to contribute to the agonist-antagonist binding site, are strongly conserved across mammalian species. Serotonin 5-HT2 receptors are widely distributed in the central nervous system and peripheral tissues. The 5-HT2 receptor class couples preferentially to Gq/G11 to increase hydrolysis of inositol phosphates and to elevate cytosolic [Ca2+]. Serotonin 5-HT2B receptor agonism may promote pulmonary and peripheral hypertension, but most importantly has been linked to serious cardiac valvulopathy. Receptor antagonism may be useful for the treatment of migraine headache.
Family:
GPCR
Sub Family:
Serotonin
Class:
Class A
Protein Name:
5-HT2B
Uniprot Number:
P41595
Protein Aliases:
5-HT2B|Serotonin receptor 2B
Gene Name:
HTR2B
Gene ID:
3357
Gene Aliases:
5-HT(2B)|5-HT2B
Accession Number:
NM_000867
Organism:
Human
Construct Details:
Full Length, Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Binding
Detection Method:
Radiometry
Measured Response:
Scintillation
Testing Information
Procedure Summary:
Human recombinant serotonin 5-HT2B receptor expressed in CHO-K1 cells are used to prepare membranes in modified Tris-HCl buffer pH 7.4. A 8 μg‡ aliquot of membrane protein is incubated with 2 nM [³H]Mesulergine for 60 minutes at 37°C. Non-specific binding is estimated in the presence of 10 μM serotonin. Membranes are filtered and washed, the filters are then counted to determine specifically bound [³H]Mesulergine. Compounds are screened at 10 μM.
Note: ‡Membrane prep properties may change from lot to lot, the concentration used will be adjusted as necessary.
Ligand:
[³H] Mesulergine
Ligand Kd (nM):
5.3
Ligand Concentration:
2 nM
Non Specific:
10 µM Serotonin (5-HT)
Incubation:
60 min at 37°C
Control Inhibitor:
SB 206553
Criteria For Significance:
≥ 50% of max stimulation or inhibition
Test Concentration / Dose:
Suggested concentration for initial testing is 10 uM
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:
Cardiovascular
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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