Neprilysin (NEP) is a transmembrane metallopeptidase normally expressed by a variety of tissues. It is also known as neutral endopeptidase, and enkephalinase. NEP cleaves peptides at the N-terminal side of hydrophobic amino acid residues and is responsible for the degradation and inactivation of a variety of physiological substrates. NEP is a major extracellular amyloid beta-peptide degrading enzyme in the brain and targeting NEP is considered a potential therapeutic strategy for the prevention and treatment of Alzheimer's disease. NEP has also been implicated in the pathogenesis of hypertension, diabetes, and cancer.
The SensoLyte® 520 Neprilysin Assay Kit employs a novel internally quenched 5-FAM/QXL™ FRET substrate for the detection of neprilysin activity. The enzyme cleaves the FRET substrate into two separate fragments resulting in the release of 5-FAM fluorescence, which can be monitored at excitation /emission= 490/520 nm. The long wavelength fluorescence of 5-FAM is less interfered by the autofluorescence of components in biological samples and test compounds. The assay can detect as low as 0.78 ng/mL of active neprilysin.