ECE1 Human Neprilysin Metallo Peptidase Enzymatic LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Endothelin (ET) is a potent vasoconstrictor that is cleaved from a low-activity precursor, big ET by endothelin converting enzymes (ECE). Two isozymes of ECE (ECE-1 and ECE-2) have been molecularly identified with each shown to belong to the type II membrane-bound metalloprotease family. ECE-1 also cleaves a variety of bioactive peptides such as bradykinin, neurotensin, angiotensin I, and substance P. An ECE-1 inhibitor may be useful for the treatment of heart failure and hypertension.
Family:
Protease
Sub Family:
Neprilysin
Class:
Metallo Peptidase
Protein Name:
ECE1
Uniprot Number:
P42892
Gene Name:
ECE1
Gene ID:
1889
Accession Number:
NM_001397
Organism:
Human
Construct Details:
Fragment (90-770), 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 ECE-1 expressed in murine myeloma cells NS0 is used. Test compound and/or vehicle is preincubated with 20 ng/ml enzyme‡ in modified MES buffer pH 6.0 for 15 minutes at 25⁰C. The reaction is initiated by addition of 10 μM Mca-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(Dnp) for another 60 minute incubation period. Determination of the amount of Mca-Arg-Pro-Pro-Gly-Phe-Ser-Ala formed is read spectrophotometrically at 320 nm/405 nm. Compounds are screened at 10 μM.
Substrate:
10 µM Mca-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(Dnp)
Incubation:
60 min at 25°C
Control Inhibitor:
Phosphoramidon
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:
Inflammation/Allergy
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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