Recombinant Mouse ECE-1 Protein, CF

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R&D Systems Recombinant Proteins and Enzymes
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Recombinant Mouse ECE-1 Protein, CF Summary

Product Specifications

>90%, 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.
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPPGFSAFK(Dnp)-OH (Catalog # ES005). The specific activity is >2,250 pmol/min/μg, as measured under the described conditions. 
Chinese Hamster Ovary cell line, CHO-derived mouse ECE-1 protein
Gln89-Trp769, with and N-terminal 6-His tag, Gln89-Trp769
Accession #
N-terminal Sequence
Structure / Form
Disulfide-linked dimer
Predicted Molecular Mass
78 kDa
90-130 kDa, reducing conditions

Product Datasheets

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Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.


Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and ZnCl2.
Shipping The product is shipped with dry ice or equivalent. 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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

  • Assay Buffer: 50 mM MES, 0.1 M NaCl, pH 6.0
  • Recombinant Human ECE-1 (rhECE-1) (Catalog # 5796-ZN)
  • Fluorogenic Peptide Substrate V: MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(DNP)-OH (Catalog # ES005), 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmECE-1 to 0.1 µg/mL in Assay Buffer.
  2. Dilute Substrate to 20 µM in Assay Buffer.
  3. Load into a black well plate 50 µL of 0.1 µg/mL of rmECE-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.
  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:
  • rmECE-1: 5 ng
  • 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: ECE-1

Endothelin-converting enzymes (ECEs) hydrolyze a specific peptide bond of big endothelins to produce active endothelins, some of the most potent vasoconstrictors known (1). ECE-1 is a member of the M13 zinc metallopeptidase family. Other members of the M13 family include thermolysin, neprilysin, Kell, and ECE-2 (2). M13 metallopeptidases can be distinguished from other metallopeptidases by their sensitivity to inhibition by phosphoramidon. ECE-1 is most highly expressed in the cardiovascular endothelium, but is also expressed in some endocrine tissues (3). ECE-1 is known to hydrolyze a variety of bioactive peptides, including bradykinin, neurotensin, angiotensins, and Substance P, with a substrate specificity similar to that of neprilysin (4). ECE-1 displays pronounced pH dependence in its substrate specificity (5). The degradation of Substance P by ECE-1 in endosomes regulates beta -arrestin-dependent ERK-2 signaling to prevent cell death in some neuronal cells (6). Four isoforms of ECE-1 are present in humans and mice, all of which encode a Type II integral membrane protein (7). The four isoforms share a common extracellular catalytic domain, differing in their N-terminal cytoplasmic tail regions. The recombinant mouse ECE-1 transmembrane and cytoplasmic tail domains were replaced with a signal sequence, resulting in the secretion of the soluble catalytic ectodomain.

  1. Yanagisawa, M. et al. (1998) Nature 332:411.
  2. Turner, A.J. et al. (2001) BioEssays 23:261.
  3. Davenport, A.P. et al. (1998) Histochem. J. 30:359.
  4. Johnson, G.D. et al. (1999) J. Biol. Chem. 274:4053.
  5. Fahnoe, D.C. et al. (2000) J. Cardiovasc. Pharmacol. 36:S22.
  6. Cottrell, G.S. et al. (2009) J. Biol. Chem. 284:22411.
  7. Lindenau, S. et al. (2006) Gene 373:109.  
Long Name
Endothelin-converting Enzyme-1
Entrez Gene IDs
1889 (Human)
Alternate Names
EC 3.4.24; EC; ECE; ECE1; ECE-1; endothelin converting enzyme 1; endothelin-converting enzyme 1


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Fluorogenic Peptide Substrates

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