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Recombinant Human Neprilysin, CF

Catalog #: 1182-ZN
5 Citations Datasheet / COA / SDS

Discontinued Product

1182-ZN has been discontinued.
View all Neprilysin/CD10 products.
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Citations (5)
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Summary Product Datasheets Carrier Free Reconstitution Calculator Background Related Research Areas

Recombinant Human Neprilysin, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPPGFSAFK(Dnp)-OH (Catalog # ES005). The specific activity is >1200 pmol/min/µg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (stably transfected)-derived human Neprilysin/CD10 protein
Tyr45-Trp743
Accession # CAA30157.1
Accession #
CAA30157.1
N-terminal Sequence
Analysis
Tyr45
Predicted Molecular Mass
80 kDa
SDS-PAGE
82 kDa, reducing conditions

Product Datasheets

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1182-ZN

Product Datasheet
COA

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

1182-ZN

Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • Assay Buffer: 50 mM Tris, 0.05% Brij-35 (v/v), pH 9.0
  • Recombinant Human Neprilysin/CD10 (rhNeprilysin) (Catalog # 1182-ZN)
  • Substrate: MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(DNP)-OH (Catalog # ES005) Prepare 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhNeprilysin to 0.1 µg/mL in Assay Buffer.
  2. Dilute Substrate to 20 µM in Assay Buffer.
  3. Load in plate 50 µL of 0.1 µg/mL of rhNeprilysin and start the reaction by adding 50 µL of 20 µM Substrate. As a Substrate Blank combine 50 µL of Substrate and 50 µL of Assay Buffer.
  4. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank

     **Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

  • rhNeprilysin: 0.005 µg
  • Substrate: 10 µM
Reconstitution Calculator

Background: Neprilysin/CD10

Neprilysin (NEP, neutral endopeptidase 24.11, EC 3.4.24.11) 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 (4, 5). NEP has also been identified as the common acute lymphoblastic leukemia antigen (CALLA), and to be expressed on the surface of lymphocytes in some disease states (2, 3). These and other observations have resulted in considerable clinical interest in NEP as a potential target for analgesics and antihypertensive drugs. NEP is also a major degrading enzyme of amyloid beta peptide (A beta ) in the brain, indicating that down-regulation of NEP activity, which could be caused by aging, can contribute to the development of Alzheimer’s disease by promoting A beta accumulation (6).

References
  1. Malfroy, B. et al. FEBS Lett. 229:206.
  2. LeTarte, M. et al. (1988) J. Exp. Med. 168:1247.
  3. Shipp, M.A. et al. (1988) Proc. Natl. Acad. Sci. USA 85:4819.
  4. Malfroy, B. et al. (1978) Nature 276:523.
  5. Kenny, A.J. and S.L. Stephenson (1988) FEBS Lett. 232:1.
  6. Itwata, N. et al. (2001) Science 292:1550.
Entrez Gene IDs
4311 (Human); 17380 (Mouse); 24590 (Rat); 101925772 (Cynomolgus Monkey)
Alternate Names
Atriopeptidase; CALLA; CALLAmembrane metallo-endopeptidase (neutral endopeptidase, enkephalinase); CD10 antigen; CD10; CD10); CD10membrane metallo-endopeptidase variant 1; Common acute lymphocytic leukemia antigen; DKFZp686O16152; EC 3.4.24; EC 3.4.24.11; Enkephalinase; EPN; Leu-19; membrane metallo-endopeptidase variant 2; membrane metallo-endopeptidase; MGC126681; MGC126707; MME; NEPmembrane metallo-endopeptidase (neutral endopeptidase, enkephalinase, CALLA; Neprilysin; Neutral Endopeptidase 24.11; Neutral endopeptidase; NKH1; SFE; Skin fibroblast elastase

Citations for Recombinant Human Neprilysin, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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  1. Immunocapture-based fluorometric assay for the measurement of neprilysin-specific enzyme activity in brain tissue homogenates and cerebrospinal fluid.
    Authors: Miners JS, Verbeek MM, Rikkert MO, Kehoe PG, Love S
    J. Neurosci. Methods, 2007-08-25;167(2):229-36.
    Species: Human
    Sample Types: Peptide
    Applications: Enzyme Assay
  2. Human Opiorphin, a natural antinociceptive modulator of opioid-dependent pathways.
    Authors: Wisner A, Dufour E, Messaoudi M, Nejdi A, Marcel A, Ungeheuer MN, Rougeot C
    Proc. Natl. Acad. Sci. U.S.A., 2006-11-13;103(47):17979-84.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  3. Using evolutionary information and ancestral sequences to understand the sequence-function relationship in GLP-1 agonists.
    Authors: Skovgaard M, Kodra JT, Gram DX, Knudsen SM, Madsen D, Liberles DA
    J. Mol. Biol., 2006-08-30;363(5):977-88.
    Species: Mouse
    Sample Types: In Vivo
    Applications: In Vivo
  4. Elevated neprilysin activity in vitreous of patients with proliferative diabetic retinopathy.
    Authors: Hara H, Oh-hashi K, Yoneda S, Shimazawa M, Inatani M, Tanihara H, Kiuchi K
    Mol. Vis., 2006-08-17;12(0):977-82.
    Species: Human
    Sample Types: Vitreous Humor
    Applications: Enzyme Assay
  5. Trafficking of nuclear heparin-binding epidermal growth factor-like growth factor into an epidermal growth factor receptor-dependent autocrine loop in response to oxidative stress.
    Authors: Kim J, Adam RM, Freeman MR
    Cancer Res., 2005-09-15;65(18):8242-9.
    Applications: Enzyme Assay

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