Neprilysin Activity Assay Kit Fluorimetric - 100 Assays
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.
Component A: 5-FAM/QXL™-520 Neprilysin substrate, Ex/Em=490/520 nm upon cleavage: 1 mM, 50 µL Component B: 5-FAM, fluorescence reference standard, Ex/Em=490/520 nm: 1 mM, 15 µL Component C: Recombinant human neprilysin: 10 µg/mL, 100 µL Component D: 2X Assay Buffer: 30 mL Component E: Inhibitor: 0.1 mM, 15 µL
UniProt number: P08473
Absorbance (nm): 490
Emission (nm): 520
Storage Conditions: Store component C at -80°C. Store all other components at -20°C. Component D can be stored at room temperature for convenience. Protect components A and B from light and moisture.
Application: FRET, Protease activity
Biomarker Target: Neprilysin
Detection Method: Fluorescent
Detection Limit: 0.78 ng/ml
Research Area: Cell & Tissue Biology, Neuroscience, Peptide & Protein Analysis
Sub-category Research Area: Alzheimer's Disease, Metabolism, Protein Assay Kits
Usage: Research use
Host: CHO (mammalian cells)
Code Nacres: NA.32