Catechol-O-Methyltransferase Human Enzymatic LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Most of the epinephrine and norepinephrine that enters the circulation, from the adrenal medulla or following administration, or that is released by exocytosis from adrenergic fibers, is methylated by catechol-o-methyltransferase (COMT) to metanephrine or normetanephrine, respectively. COMT (EC 2.1.2.6) is a soluble magnesium-requiring cytoplasmic enzyme that plays a major role in the metabolism of endogenous circulating and administered catecholamine.
Family:
Neurotransmitter Metabolism
Sub Family:
Catecholamine Turnover
Protein Name:
Catechol-O-Methyltransferase
Uniprot Number:
P21964
Gene Name:
COMT
Gene ID:
1312
Accession Number:
NM_000754
Organism:
Human
Construct Details:
Full Length, Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Spectrofluorimetry
Measured Response:
Fluorescence
Testing Information
Procedure Summary:
Human recombinant COMT expressed in Sf21 insect cells is used. Test compound and/or vehicle are preincubated with 19.5 μg/mlǂ of enzyme for 15 minutes at 37°C in Tris buffer pH 8.0. The reaction is initiated by addition of 10 μM Esculetin for another 30 minute incubation period. Read plate on a spectrophotometer (Excitation: 340 nm, Emission: 460 nm). Compounds are screened at 10 μM.
Substrate:
10 µM Esculetin
Incubation:
30 min at 37°C
Control Inhibitor:
Ro 41-0960
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:
CNS/Addiction/Pain
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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