Recombinant Human Neprilysin-2/MMEL1 Protein, CF

R&D Systems | Catalog # 2340-ZN

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Neprilysin-2/MMEL1 Protein (2340-ZN)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

AAL08942

Applications

Enzyme Activity
View all Neprilysin-2/MMEL1 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Neprilysin-2/MMEL1 protein
Gly69-Trp770, with an N-terminal 6-His tag

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.

N-terminal Sequence Analysis

His

Predicted Molecular Mass

82 kDa

SDS-PAGE

99 kDa, reducing conditions

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.

Formulation, Preparation, and Storage

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.

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.

Alternate Names

MMEL1, MMEL2, Neprilysin2, NL2, SEP

Entrez Gene IDs

79258 (Human)

Gene Symbol

MMEL1

UniProt

AAL08942

Additional Neprilysin-2/MMEL1 Products

Product Documents for Recombinant Human Neprilysin-2/MMEL1 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Neprilysin-2/MMEL1 Protein, CF

For research use only

Related Research Areas

Metalloproteases and Regulators

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

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Protocols

View specific protocols for Recombinant Human Neprilysin-2/MMEL1 Protein, CF (2340-ZN):

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

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