Recombinant Rat ACE-2 Protein, CF

Catalog #: 4516-ZN Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
4516-ZN-010

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Recombinant Rat ACE-2 Protein, CF Summary

Product Specifications

Purity

>95%, 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.

Activity

Measured by its ability to cleave a fluorogenic peptide substrate, Mca-YVADAPK(Dnp)-OH (Catalog # ES007). The specific activity is >300 pmol/min/µg, as measured under the described conditions.

Source

Mouse myeloma cell line, NS0-derived rat ACE-2 protein
Met1-Thr740, with a C-terminal 10-His tag

Accession #

NP_001012006

N-terminal Sequence
Analysis

No results obtained: Gln18 predicted

Predicted Molecular Mass

85 kDa

SDS-PAGE

120 kDa, reducing conditions

Product Datasheets

4516-ZN

Product Datasheet

COA

SDS

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.

4516-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.

Assay Procedure

Materials

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 V_max* (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:

rrACE-2: 0.025 µg
Substrate: 10 µM

Reconstitution Calculator

Background: ACE-2

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).

References

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.

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