ECEs or Endothelin converting enzymes (ECE-1/ECE-2) are known as rate-limiting enzymes for the formation of the vasoconstrictor peptide endothelin-1 (ET-1) from its precursor proendothelin-1. ECE-1 can also degrade amyloid ß fragments both in vitro and in vivo. Highly homologous and structurally comparable to ECE-1, ECE-2 was shown in homozygous knockout mice to have decreased Aß degradation and significant elevated Aß 1-40 and Aß 1-42 in the brain. The SensoLyte® 520 ECEs Activity Assay Kit can be used to detect ECE activity in biological samples or in purified enzyme preparations. The substrate provided in this kit was based on a beta amyloid fragment. When active ECEs cleave the FRET substrate, it results in an increase of 5-FAM fluorescence, and is monitored at excitation/emission = 490/520 nm. The long wavelength fluorescence of 5-FAM shows less interference from autofluorescence of components in biological samples and test compounds. This assay can detect as low as 15.6 ng/mL active ECE-1 and 62.5 ng/mL active ECE-2.
The kit contains:
5-FAM/QXL® 520 ECEs substrate, Ex/Em=490/520 nm upon cleavage
Assay buffer
Human recombinant ECE-1
Inhibitor
Fluorescence reference standard for calibration
Detailed protocol
Kit size: 100 assays (96-well plate not included)
Benefits:
Industry's First available kit for ECEs Activity
Long wavelength fluorimetric kit means lower background, higher sensitivity
Datasheet
Material Safety Data Sheets (MSDS)
