Recombinant Rat ACE-2 Protein, CF
Recombinant Rat ACE-2 Protein, CF Summary
Met1-Thr740, with a C-terminal 10-His tag
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.
|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.
- Assay Buffer: 50 mM Tris, 1 M NaCl, pH 7.5
- Recombinant Rat ACE-2 (rrACE-2) (Catalog # 4516-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 rrACE-2 to 0.5 ng/μL in Assay Buffer.
- Dilute Substrate 20 μM in Assay Buffer.
- In plate load 50 μL of 0.5 ng/μL rrACE-2. In a separate well load 50 µL Assay Buffer to be used as a Substrate Blank.
- Start the reaction by adding 50 μL of 20 μM Substrate to all wells.
- 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).
- rrACE-2: 0.025 µg
- Substrate: 10 µM
Angiotensin I Converting Enzyme 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). The predicted rat ACE-2 protein sequence consists of 805 amino acids, including an 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 cardiovascular function in vivo (3). In addition, ACE-2 has a critical function in acute lung injury (4).
- 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. (2006) Curr. Opinion Pharmacol. 6:271.
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Fluorogenic Peptide Substrates
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