Recombinant Mouse ACE-2 Protein, CF

Catalog # Availability Size / Price Qty
3437-ZN-010
Recombinant Mouse ACE-2 Protein Enzyme Activity
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Citations (3)
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Recombinant Mouse ACE-2 Protein, CF Summary

Product Specifications

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.
Activity
Measured by its ability to cleave a fluorogenic peptide substrate, Mca-YVADAPK(Dnp)-OH (Catalog # ES007). The specific activity is >400 pmol/min/µg, as measured under the described conditions.
Source
Chinese Hamster Ovary cell line, CHO-derived mouse ACE-2 protein
Gln18-Thr740, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
No results obtained: Gln18 predicted
Structure / Form
Recombinant Mouse ACE‑2 is prone to proteolytic cleavage at C-terminus. The predominant form of the purified protein lacks the His tag.
Predicted Molecular Mass
85 kDa
SDS-PAGE
110 kDa, reducing conditions

Product Datasheets

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3437-ZN

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.

3437-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, 1 M NaCl, pH 7.5
  • Recombinant Mouse ACE-2 (rmACE-2) (Catalog # 3437-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)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmACE-2 to 0.5 ng/µL in Assay Buffer.
  2. Dilute Substrate to 20 µM in Assay Buffer.
  3. In a plate load 50 µL of 0.5 ng/µL rmACE-2, and start the reaction by adding 50 µL of 20 µM Substrate to wells. 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:
  • rmACE-2: 0.025 µg
  • Substrate: 10 µM

























Scientific Data

Enzyme Activity Recombinant Mouse ACE-2 Protein Enzyme Activity View Larger

Recombinant Mouse ACE-2 His-tag (Catalog # 3437-ZN) is measured by its ability to cleave a fluorogenic peptide substrate, Mca-YVADAPK(Dnp)-OH (Catalog # ES007).

Reconstitution Calculator

Reconstitution Calculator

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Background: ACE-2

ACE-2, also called ACEH (ACE homologue), is an integral membrane protein and a zinc metalloprotease of the ACE family that also includes somatic and germinal ACE (1). Mouse ACE-2 has about 40% amino acid identity to the N- and C-terminal domains of mouse somatic ACE. The predicted mouse ACE-2 protein sequence consists of 798 amino acids, including a N-terminal signal peptide, a single catalytic domain, a C-terminal membrane anchor, and a short cytoplasmic tail. ACE-2 cleaves angiotensins I and II as a carboxypeptidase. ACE-2 mRNA is found at high levels in testis, kidney and heart and at moderate levels in colon, small intestine and ovary. Classical ACE inhibitors such as captopril and lisinopril do not inhibit ACE-2 activity. Novel peptide inhibitors of ACE-2 do not inhibit ACE activity (2). Genetic data from Drosophila, mice and rats show that ACE-2 is an essential regulator of heart function in vivo (3). In addition, ACE-2 is a key SARS-CoV Spike protein receptor in vivo and has a critical function in acute lung injury (4, 5).

References
  1. Tipnis, S.R. et al. (2000) J. Biol. Chem. 275:33238.
  2. Crackower, M.A. et al. (2002) Nature 417:822.
  3. Huang, L. et al. (2003) J. Biol. Chem. 278:15532.
  4. Kuba, K. et al. (2005) Nature Med. 11:875.
  5. Ima, Y. et al. (2005) Nature 436:112.
Long Name
Angiotensin I Converting Enzyme 2
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)
Alternate Names
ACE2; ACE-2; ACEH; ACEHangiotensin I converting enzyme 2; ACE-related carboxypeptidase; angiotensin I converting enzyme (peptidyl-dipeptidase A) 2; angiotensin-converting enzyme 2; Angiotensin-converting enzyme homolog; DKFZp434A014; EC 3.4.17; EC 3.4.17.23; Metalloprotease MPROT15

Citations for Recombinant Mouse ACE-2 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.

3 Citations: Showing 1 - 3
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  1. ACE2 is augmented in dystrophic skeletal muscle and plays a role in decreasing associated fibrosis.
    Authors: Riquelme C, Acuna M, Torrejon J, Rebolledo D, Cabrera D, Santos R, Brandan E
    PLoS ONE, 2014-04-02;9(4):e93449.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Bioassay
  2. ACE2 links amino acid malnutrition to microbial ecology and intestinal inflammation.
    Authors: Hashimoto T, Perlot T, Rehman A
    Nature, 2012-07-25;487(7408):477-81.
    Species: Mouse
    Sample Types: In Vivo
    Applications: In Vivo
  3. Major role for ACE-independent intrarenal ANG II formation in type II diabetes.
    Authors: Park S, Bivona BJ, Kobori H, Seth DM, Chappell MC, Lazartigues E, Harrison-Bernard LM
    Am. J. Physiol. Renal Physiol., 2009-10-21;298(1):F37-48.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Enzyme Assay

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