Active Neprilysin (CD10)

NEP; MME; CALLA; SFE; Atriopeptidase; Enkephalinase; Skin fibroblast elastase; Common Acute Lymphoblastic Leukemia Antigen; Membrane Metallo-Endopeptidase

ACTIVITY TEST

Neprilysin/CD10, also known as NEP and neutral endopeptidase, is a zinc metallopeptidase expressed at the cell surface of a variety of cells. The enzyme functions both as an endopeptidase with a thermolysin-like specificity and as a dipeptidylcarboxypeptidase. NEP has been shown to be involved in the degradation of enkephalins in the mammalian brain and the inactivation of circulating atrial natriuretic peptide. NEP has also been identified as the common acute lymphocytic leukemia antigen (CALLA), and is expressed on the surface of lymphocytes in some disease states. The activity of recombinant human NEP is measured by its ability to cleave a fluorogenic peptide substrate MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(DNP)-OH in the assay buffer 50 mM Tris, 0.05% Brij-35, pH 9.0. The rhNEP is diluted to 0.1 ug/ml in assay buffer, then add 50 µL protein into a black well plate and start the reaction by adding 50 µL of 20 µM substrate (diluted by assay buffer), with a substrate blank containing 50 µL assay buffer, 50 µL substrate, and no rhNEP. Then read at excitiation and emission wavelengths of 320 nm and 405 nm, respectively, in kinetic mode for 5 minutes. The specific activity of recombinant human NEP is > 3500 pmol/min/µg.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
13 Elevated Plasma levels of Neutral Endpeptidase in Hypertension Patients Treated with Angiotensin Converting Enzyme Inhibitors Ahajournals:Source
Biol Pharm Bull Effect of High Glucose Levels on Amyloid β Production in Retinas of Spontaneous Diabetes Mellitus Otsuka Long-Evans Tokushima Fatty Rats PubMed: 25832640
Eur J Heart Fail.  Soluble neprilysin does not correlate with outcome in heart failure with preserved ejection fraction Pubmed:26725876
8 A Test in Context: Neprilysin: Function, Inhibition, and Biomarker. pubmed:27491909
European Journal of Heart Failure Effects of sacubitril/valsartan on neprilysin targets and the metabolism of natriuretic peptides in chronic heart failure: a mechanistic clinical study Pubmed: 30520545
European Journal of Heart Failure Increased granulocyte membrane neprilysin (CD10) expression is associated with better prognosis in heart failure Pubmed: 30828920
Clinical Research in Cardiology Soluble neprilysin, NT-proBNP, and growth differentiation factor-15 as biomarkers for heart failure in dialysis patients (SONGBIRD) Pubmed: 32002632
Neurotoxicology Involvement of hippocampal agmatine in β1-42 amyloid induced memory impairment, neuroinflammation and BDNF signaling disruption in mice Pubmed: 32522471
European Heart Journal P1620 Association of granulocyte neprilysin (CD10) expression with prognosis in heart failure with reduced ejection fraction
Clin Biochem Finding a reliable assay for soluble neprilysin Pubmed:35331754
Scientific Reports The potential role of serum tau protein (MAPT), neuronal cell adhesion molecule (NrCAM) and neprilysin (NEP) in neurodegenerative disorders development in …
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