Recombinant Human ECE-1 Protein, CF Summary
Product Specifications
Gln90-Trp770, with an N-terminal signal peptide, 10-His tag and Val
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.
1784-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: 0.1 M MES, 0.1 M NaCl, pH 6.0
- Recombinant Human ECE-1 (rhECE-1) (Catalog # 1784-ZN)
- Fluorogenic Peptide Substrate V: 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 rhECE-1 to 0.1 µg/mL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load into a black well plate 50 µL of 0.1 µg/mL of rhECE-1, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL 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).
- rhECE-1: 0.005 μg
- Substrate: 10 µM
Reconstitution Calculator
Background: ECE-1
Endothelin-converting Enzyme 1 (ECE-1) is a zinc protease of the neprilysin (NEP) family, which also includes ECE-2, PEX, XCE, DINE, Kell and several NEP-like proteins (1). ECE-1 is a type II transmembrane protein with a short cytoplasmic tail and a large ectodomain. Four alternatively spliced isoforms differ in their cytoplasmic tail (2, 3). In addition to big endothelin-1, ECE-1 cleaves a variety of bioactive peptides such as bradykinin, neurotensin, angiotensin I, and substance P (1). Together with ECE-2, it is also involved in degradation of beta -amyloid peptide (4). The ectodomain of human ECE-1, which is common to all isoforms, was expressed with an N-terminal His tag and purified.
- Turner, A.J. et al. (2001) BioEssays 23:261.
- Valdennaire, O. et al. (1999) Eur. J. Biochem. 264:341.
- Schweizer, A. et al. (1997) Biochem. J. 328:871.
- Eckman, E.A. et al. (2003) J. Biol. Chem. 278:2081.
Citations for Recombinant Human ECE-1 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.
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Citations: Showing 1 - 7
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Ligand-Specific Factors Influencing GLP-1 Receptor Post-Endocytic Trafficking and Degradation in Pancreatic Beta Cells
Authors: Z Fang, S Chen, Y Manchanda, S Bitsi, P Pickford, A David, MM Shchepinov, IR Corrêa, DJ Hodson, J Broichhage, EW Tate, F Reimann, V Salem, GA Rutter, T Tan, SR Bloom, A Tomas, B Jones
Int J Mol Sci, 2020-11-09;21(21):.
Species: Human
Sample Types: Protein
Applications: Bioassay -
A stress-enhanced model for discovery of disease-modifying gene: Ece1-suppresses the toxicity of ?-synuclein A30P
Authors: AY Chen, T Tully
Neurobiol. Dis., 2018-03-07;114(0):153-163.
Species: Human
Sample Types: Fluorogenic Peptide Substrate
Applications: Bioassay -
Endothelin-converting enzymes degrade ?-synuclein and are reduced in dementia with Lewy bodies
Authors: JS Miners, S Love
J. Neurochem, 2017-03-29;0(0):.
Applications: Bioassay -
Endothelin-converting enzyme-2 is increased in Alzheimer's disease and up-regulated by Abeta.
Authors: Palmer JC, Baig S, Kehoe PG, Love S
Am. J. Pathol., 2009-06-18;175(1):262-70.
Applications: Western Blot -
Endothelin-converting enzyme-1 degrades internalized somatostatin-14.
Authors: Roosterman D, Kempkes C, Cottrell GS, Padilla BE, Bunnett NW, Turck CW, Steinhoff M
Endocrinology, 2008-02-14;149(5):2200-7.
Species: Human
Sample Types: Recombinant Protein
Applications: Enzyme Assay -
Endothelin-converting enzyme-1 regulates endosomal sorting of calcitonin receptor-like receptor and beta-arrestins.
Authors: Padilla BE, Cottrell GS, Roosterman D, Pikios S, Muller L, Steinhoff M, Bunnett NW
J. Cell Biol., 2007-11-26;179(5):981-97.
Applications: Enzyme Assay -
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
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