OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
APJ (apelin) belongs to the superfamily of seven transmembrane G-protein-coupled receptors, which 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. APJ receptors are found in the lung, heart, and hypothalamic region.
Family:
GPCR
Sub Family:
Apelin
Class:
Class A
Protein Name:
APJ
Uniprot Number:
P35414
Protein Aliases:
APJ (apelin) receptor
Gene Name:
APLNR
Gene ID:
187
Gene Aliases:
FLJ90771|APJ|APJR
Accession Number:
NM_005161
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:
Chem-1 cells stably transfected with a plasmid encoding human APJ receptors are used to prepare membranes in modified HEPES buffer pH 7.4. A 0.1 µg‡ aliquot of membrane is incubated with 0.05 nM [¹²⁵I] (Glp⁶⁵, Nle⁷⁵, Tyr⁷⁷)-Apelin-¹³ for 120 minutes at 25°C. Non-specific binding is estimated in the presence of 1 µM (Pyr1)-Apelin-13. Membranes are filtered and washed, the filters are then counted to determine [¹²⁵I] (Glp⁶⁵, Nle⁷⁵, Tyr⁷⁷)-Apelin-¹³ specifically bound. Compounds are screened at 10 µM.
Note: ‡Membrane protein may change from lot to lot, the concentration used will be adjusted if necessary.
Ligand:
[¹²⁵I] (Glp⁶⁵, Nle⁷⁵, Tyr⁷⁷)-Apelin-¹³
Ligand Kd (nM):
0.05
Ligand Concentration:
0.05 nM
Non Specific:
1 µM (Pyr¹)-Apelin-13
Incubation:
120 min at 25°C
Control Inhibitor:
(Pyr¹)-Apelin-13
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:
Metabolic Diseases
Adverse / Beneficial Action:
APJ receptors agonism may cause hypotension and protection against HIV cell entry.
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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