Recombinant Human Neprilysin-2/MMEL1 Protein, CF

  (1 citations)     
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Citations (1)
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  • 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

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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
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
Related Research Areas
Citations:

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 Citations: 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 Type: Recombinant Protein
    Application: EnzAct

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Fluorogenic Peptide Substrates
Description Application Cat# Citations Images  

Mca-RPPGFSAFK(Dnp)-OH Fluorogenic Peptide Substrate

BA , BA , Bind ES005 9
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Primary Antibodies
Description Application Cat# Citations Images  

His Tag Antibody

WB , Simple Western , Flow , AP , CyTOF-ready MAB050 28  
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His Tag HRP-conjugated Antibody

WB , Simple Western MAB050H 3  
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His Tag APC-conjugated Antibody

ICFlow IC050A  
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His Tag Biotinylated Antibody

WB BAM050 4  
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His Tag PE-conjugated Antibody

ICFlow IC050P 3  
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His Tag Antibody

WB , Flow , AP MAB050R 2  
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His Tag Alexa Fluor® 488-conjugated Antibody

ICFlow IC050G  
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His Tag PerCP-conjugated Antibody

ICFlow IC050C  
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His Tag Alexa Fluor® 700-conjugated Antibody

ICFlow IC050N  
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Protein Purification Kits
Description Application Cat# Citations Images  

Histidine-tagged Protein Purification Resin

IP999 1
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