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

PubChem ID

3822

Name

Ketanserin

Measurement

0.34μM - IC50

PubChem ID

5163

Name

SB 206553

Measurement

0.016μM - IC50

PubChem ID

68848

Name

Mesulergine

Measurement

0.0058μM - IC50

PubChem ID

3277600

Name

SB 204741

Measurement

0.24μM - IC50

PubChem ID

24278124

Name

Serotonin (5-HT)

Measurement

0.057μM - IC50

PubChem ID

Name

(δ)-DOI

Measurement

0.15μM - IC50

Clinical Relevance

Therapeutic Area:

Cardiovascular

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei

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