Recombinant Human ACE-2 Fc Chimera Avi-tag Protein (CHO), CF

Biotinylated
Catalog # Availability Size / Price Qty
AVI10544-020
Recombinant Human ACE-2 Fc Chimera Avi-tag Protein (CHO), CF Binding Activity
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Recombinant Human ACE-2 Fc Chimera Avi-tag Protein (CHO), CF Summary

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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 binding ability in a functional ELISA. When Recombinant SARS-CoV-2 Spike RBD Fc Chimera (Catalog # 10499-CV) is immobilized at 0.2 µg/mL (100 µL/well), Biotinylated Recombinant Human ACE-2 Fc Chimera Avi-tag (CHO Expressed) (Catalog # AVI10544) binds with an ED50 of 0.6-4.8. Measured by its ability to cleave a fluorogenic peptide substrate, Mca-YVADAPK(Dnp)-OH (Catalog # ES007). The specific activity is >500 pmol/min/μg, as measured under the described conditions.
Source
Chinese Hamster Ovary cell line, CHO-derived human ACE-2 protein
Human ACE-2
(Gln18-Ser740)
Accession # Q9BYF1.2
IEGRMDHuman IgG1
(Pro100-Lys330)
Avi-tag
N-terminusC-terminus
Accession #
N-terminal Sequence
Analysis
Protein identity confirmed by mass spectrometry.
Structure / Form
Biotinylated via Avi-tag
Predicted Molecular Mass
112.0 kDa
SDS-PAGE
118-130 kDa, under reducing conditions

Product Datasheets

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.

AVI10544

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.

Data Images

Binding Activity View Larger

When Recombinant SARS-CoV-2 Spike RBD Fc Chimera (10499-CV) is immobilized at 0.2 µg/mL (100 µL/well), Biotinylated Recombinant Human ACE-2 Fc Chimera Avi-tag (CHO Expressed) (Catalog # AVI10544) binds with an ED50 of 0.6-4.8 ng/mL.

Enzyme Activity View Larger

Biotinylated Recombinant Human ACE‑2 Fc Chimera Avi-tag (CHO Expressed) (Catalog # AVI10544) is measured by its ability to cleave a fluorogenic peptide substrate, Mca-YVADAPK(Dnp)-OH (ES007).

SDS-PAGE View Larger

2 μg/lane of Biotinylated Recombinant Human ACE-2 Fc Chimera Avi-tag Protein (Catalog # AVI10544) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at ~124 kDa under reducing conditions.

Reconstitution Calculator

Reconstitution Calculator

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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.181:1.
  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
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

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