Recombinant Human Neprilysin-2/MMEL1 Protein, CF

Catalog # Availability Size / Price Qty
2340-ZN-010
Product Details
Citations (1)
FAQs
Supplemental Products
Reviews

Recombinant Human Neprilysin-2/MMEL1 Protein, CF Summary

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 >250 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Neprilysin-2/MMEL1 protein
Gly69-Trp770, with an N-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
His
Predicted Molecular Mass
82 kDa
SDS-PAGE
99 kDa, reducing conditions

Product Datasheets

2340-ZN

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and ZnCl2.
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, 250 mM NaCl, pH 7.5
  • Recombinant Human Neprilysin-2/MMEL1 (rhNeprilysin-2) (Catalog # 2340-ZN)
  • Substrate: MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(DNP)-OH (Catalog # ES005)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhNeprilysin-2 to 1 µg/mL in Assay Buffer.
  2. Dilute Substrate to 20 µM in Assay Buffer.
  3. Load into a black well plate 50 µL of 1 µg/mL rhNeprilysin-2, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 20 µM Substrate.
  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).

Per Well:
  • rhNeprilysin-2: 0.050 µg
  • Substrate: 10 µM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

=
÷

Background: Neprilysin-2/MMEL1

Neprilysin-2 is a zinc protease of the neprilysin (NEP) family, which also includes NEP, ECE-2, PEX, DINE, Kell and several NEP-like proteins (1). It is encoded by the MMEL1 gene in the human genome and is also known as soluble secreted endopeptidase (SEP), Neprilysin-like 1 and Neprilysin-2 (2). Highly expressed in testis, the cDNA predicted a type II transmembrane protein with a short cytoplasmic tail and a large ectodomain. Both membrane-bound and soluble forms were comparable with regard to model substrates, pH optima and inhibitor profiles. The ectodomain of human Neprilysin-2/MMEL1 was expressed with an N-terminal His tag and purified.

References
  1. Turner, A.J. et al. (2001) BioEssays 23:261.
  2. Carpentier, M. et al. (2004) in Handbook of Proteolytic Enzymes. Barrett, A.J. et al. eds. p. 426.
Entrez Gene IDs
79258 (Human)
Alternate Names
EC 3.4.24; EC 3.4.24.11; MELL1; membrane metallo-endopeptidase-like 1; Membrane metallo-endopeptidase-like 2MGC119455; MGC119454; MMEL1; MMEL2; NEP2; NEP2(m); NEPIIMGC119456; Neprilysin II; Neprilysin2; Neprilysin-2; NL2; NL2NL1; SEP; soluble secreted endopeptidase; zinc metallopeptidase

Citation for Recombinant Human Neprilysin-2/MMEL1 Protein, 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.

1 Citation: Showing 1 - 1

  1. Human membrane metallo-endopeptidase-like protein degrades both beta-amyloid 42 and beta-amyloid 40.
    Authors: Huang JY, Bruno AM, Patel CA, Huynh AM, Philibert KD, Glucksman MJ, Marr RA
    Neuroscience, 2008;155(1):258-62.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: EnzAct

FAQs

No product specific FAQs exist for this product, however you may

View all Proteins and Enzyme FAQs

Reviews for Recombinant Human Neprilysin-2/MMEL1 Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human Neprilysin-2/MMEL1 Protein, CF and earn rewards!

Have you used Recombinant Human Neprilysin-2/MMEL1 Protein, CF?

Submit a review and receive an Amazon gift card.

$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image

$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image

Submit a Review