Recombinant Human Neprilysin-2/MMEL1 Protein, CF
Recombinant Human Neprilysin-2/MMEL1 Protein, CF Summary
Product Specifications
Gly69-Trp770, with an N-terminal 6-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
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.
|
Assay Procedure
- 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
- Dilute rhNeprilysin-2 to 1 µg/mL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- 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.
- Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
- 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-2: 0.050 µg
- Substrate: 10 µM
Reconstitution Calculator
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.
- Turner, A.J. et al. (2001) BioEssays 23:261.
- Carpentier, M. et al. (2004) in Handbook of Proteolytic Enzymes. Barrett, A.J. et al. eds. p. 426.
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
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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, 2008-05-16;155(1):258-62.
Species: Human
Sample Types: N/A, Recombinant Protein
Applications: Enzyme Assay, Western Blot
FAQs
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