Recombinant Mouse ACE-2 Protein, CF Summary
Product Specifications
Gln18-Thr740, with a C-terminal 10-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.
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.
|
Assay Procedure
- 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
- Dilute rmACE-2 to 0.5 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- 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.
- 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).
Per Well:- rmACE-2: 0.025 µg
- Substrate: 10 µM
Scientific Data
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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
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).
- Tipnis, S.R. et al. (2000) J. Biol. Chem. 275:33238.
- Crackower, M.A. et al. (2002) Nature 417:822.
- Huang, L. et al. (2003) J. Biol. Chem. 278:15532.
- Kuba, K. et al. (2005) Nature Med. 11:875.
- Ima, Y. et al. (2005) Nature 436:112.
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.
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Citations: Showing 1 - 3
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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 -
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 -
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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