Recombinant Ferret ACE-2 His-tag Protein, CF

R&D Systems | Catalog # 10635-ZN

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Key Product Details

  • R&D Systems HEK293-derived Recombinant Ferret ACE-2 His-tag Protein (10635-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

HEK293

Accession Number

NP_001297119.1

Applications

Enzyme Activity
View all ACE-2 Proteins and Enzymes »

Product Specifications

Source

Human embryonic kidney cell, HEK293-derived ferret ACE-2 protein
Gln18-Thr740
with a C-terminal 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Protein identity confirmed by mass spectrometry

Predicted Molecular Mass

85 kDa

SDS-PAGE

90-105 kDa, under reducing conditions

Activity

Measured by its ability to cleave a fluorogenic peptide substrate, Mca-YVADAPK(Dnp)-OH (Catalog # ES007).
The specific activity is >100 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Ferret ACE-2 His-tag Protein, CF

Recombinant Ferret ACE-2 His-tag Protein Enzyme Activity.

Recombinant Ferret ACE‑2 His-tag Protein (Catalog # 10635-ZN) is measured by its ability to cleave a fluorogenic peptide substrate, Mca-YVADAPK(Dnp)-OH (ES007).

Recombinant Ferret ACE-2 His-tag Protein SDS-PAGE.

2 μg/lane of Recombinant Ferret ACE-2 His-tag (Catalog # 10635-ZN) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at ~101 kDa under reducing conditions.

Formulation, Preparation, and Storage

10635-ZN
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, ZnCl2 and Glycerol.
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: ACE-2

Angiotensin I Converting Enzyme (ACE-2), also called ACEH (ACE homologue), is a dimeric, zinc-dependent metalloprotease of the ACE family that also includes somatic and germinal ACE (1, 2). ACE-2 mRNA is found at high levels in heart, testis, and kidney and at lower levels in a wide variety of tissues (1, 3). ACE-2 is the SARS-CoV and SARS-CoV2 Spike protein receptor in vivo (4-6), functions catalytically as a carboxypeptidase to cleave several substrates including angiotensins I and II, and acts as a partner for B0AT1-family amino acid transporters (1, 2). Through these functions, ACE-2 has been shown to be involved in several diseases including SARS, COVID19, acute lung injury (4, 7), heart disease (8), liver and lung fibrosis (9), inflammatory lung disease (10), and cardiopulmonary disease (11). Full length ACE-2 protein includes an extracellular region composed of a single N-terminal peptidase domain and C-terminal collectrin-like domain (CLD), a transmembrane domain, and a short cytoplasmic tail (12). The N-terminal peptidase region is required for binding to SARS-CoV and SARSCoV2 spike proteins, while the CLD contains a region that promotes dimerization and association with amino acid transporters (2). The peptidase domain contains a long deep cleft that undergoes a large hinge-bending movement at substrate and inhibitor binding (12).  Classical ACE inhibitors such as captopril and lisinopril do not inhibit ACE-2 activity and inhibitors of ACE-2 do not inhibit ACE activity (13).    

References

  1. Kuba, K. et al. (2010) Pharmacol. Ther. 128:119.
  2. Yan, R. et al. (2020) Science 367:1444.
  3. Tipnis, S.R. et al. (2000) J. Biol. Chem. 275:33238.
  4. Kuba, K. et al. (2005) Nature Med. 11:875.
  5. Hoffmann, M. et al. (2020) Cell 181:271.
  6. Wrapp, D. et al. (2020) Science 367:1260.
  7. Imai, Y. et al. (2005) Nature 436:112.
  8. Huang, L. et al. (2003) J. Biol. Chem. 278:15532.
  9. Schrom, E. et al. (2017) Mol. Therapy Nuc. Acid 7:350.
  10. Jia, H. et al. (2016) Shock. 46:239.
  11. Cole-Jeffrey, C.T. et al. (2015) J. Cardiovasc. Pharmacol. 66:540.
  12. Towler, P. et al. (2004) J. Biol. Chem. 279:17996.
  13. Crackower, M.A. et al. (2002) Nature 417:822.

Long Name

Angiotensin I Converting Enzyme 2

Alternate Names

ACE2, ACEH

Entrez Gene IDs

59272 (Human); 70008 (Mouse); 302668 (Rat); 100144303 (Porcine); 480847 (Canine); 418623 (Chicken); 102130864 (Cynomolgus Monkey); 554349 (Feline); 101673097 (Ferret); 101823817 (Hamster); 108390919 (Malayan Pangolin)

Gene Symbol

ACE2

UniProt

NP_001297119.1

Additional ACE-2 Products

Product Documents for Recombinant Ferret ACE-2 His-tag Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Ferret ACE-2 His-tag Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Ferret ACE-2 His-tag Protein, CF (10635-ZN):

Materials
  • Assay Buffer: 75 mM Tris, 1 M NaCl, pH 7.5
  • Recombinant Ferret ACE-2 (rferretACE-2) (Catalog # 10635-ZN)
  • Substrate:  Mca-Tyr-Val-Ala-Asp-Ala-Pro-Lys(Dnp)-OH (Catalog # ES007), 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rferretACE-2 to 0.8 µg/mL in Assay Buffer.
  2. Dilute Substrate to 60 µM in Assay Buffer.
  3. Load into a plate 50 µL of 0.8 µg/mL rferretACE-2, and start the reaction by adding 50 µL of 60 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 60 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:

  • rferretACE-2: 0.04 µg
  • Substrate: 30 µM

FAQs

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