Recombinant Human ACE-2 Protein, CF

Name: Recombinant Human ACE-2 Protein, CF
Brand: R&D Systems
SKU: 933-ZN
Rating: 5 (5 reviews)

R&D Systems, Inc. a Bio-Techne Brand

Catalog # 933-ZN

Citations (24)

(5)

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Key Product Details

Source:	NS0
Accession #:	Q9BYF1
Structure / Form:	Recombinant Human ACE‑2 is prone to proteolytic cleavage at C-terminus.
Applications:	Bioactivity, Enzyme Activity

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Product Specifications for Recombinant Human ACE-2 Protein, CF

Source	Mouse myeloma cell line, NS0-derived human ACE-2 protein Gln18-Ser740, with a C-terminal 10-His tag
Purity	>90%, 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.
N-terminal sequence Analysis	No results obtained: Gln18 predicted
Predicted Molecular Mass	85 kDa
SDS-PAGE	101-111 kDa, reducing conditions
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), Recombinant Human ACE-2 (Catalog # 933-ZN) binds with an ED₅₀ of 1.00-12.0 ng/mL. Measured by its ability to cleave a fluorogenic peptide substrate, Mca-YVADAPK(Dnp)-OH (Catalog # ES007). The specific activity is >800 pmol/min/µg, as measured under the described conditions.

Scientific Data Examples for Recombinant Human ACE-2 Protein, CF

	Recombinant Human ACE‑2 Protein SEC-MALS. Recombinant Human ACE-2/His (Catalog # 933-ZN) has a molecular weight (MW) of 189.9 kDa as analyzed by SEC-MALS, suggesting that this protein is a homodimer. MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).
	Multiple Lots of Recombinant Human ACE-2 bind SARS-CoV-2 Spike RBD Protein Four independent lots of recombinant Human ACE-2 His-tag 933-ZN were tested in a functional ELISA for binding to recombinant SARS-CoV-2 Spike RBD Fc Chimera Protein 10499-CV. All four lots were plotted on the same graph to show lot-to-lot consistency of recombinant Human ACE-2. The ED₅₀ for this effect is 1.00-12.0 ng/mL.
	SPR Data of Recombinant Human ACE-2 binding to SARS-CoV-2 Spike RBD protein Recombinant SARS-CoV-2 Spike RBD Fc Chimera Protein (10499-CV) was immobilized on a Biacore Sensor Chip CM5, and binding to recombinant human ACE-2 (933-ZN) was measured at a concentration range between 0.73 nM and 46.7 nM. The double-referenced sensorgram was fit to a 1:1 binding model to determine the binding kinetics and affinity, with an affinity constant of K_d=7.037 nM. (Biacore T200).
	Multiple Lots of Recombinant Human ACE-2 bind SARS-CoV-2 Spike RBD Protein Recombinant Human ACE-2 His-tag (Catalog # 933-ZN) is measured by its ability to cleave fluorogenic peptide substrate, Mca-YVADAPK(Dnp)-OH (ES007).
	1 μg/lane of Recombinant Human ACE-2 Protein (Catalog # 933-ZN) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing bands at ~106 kDa under reducing conditions.

Formulation, Preparation and Storage

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.

933-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.

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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).

Item	Value
References	Kuba, K. et al. (2010) Pharmacol. Ther. 128:119. Yan, et al. (2020) Science 367:1444. Tipnis, S.R. et al. (2000) J. Biol. Chem. 275:33238. Kuba, K. et al. (2005) Nature Med. 11:875. Hoffmann, M. et al. (2020) Cell.181:1. Wrapp, et al. (2020) Science 367:1260. Imai, Y. et al. (2005) Nature 436:112. Huang, L. et al. (2003) J. Biol. Chem. 278:15532. Schrom, E. et al. (2017) Mol. Therapy Nuc. Acid 7:350. Jia, H. et al. (2016) Shock. 46:239. Cole-Jeffrey, C.T. et al. (2015) J. Cadiovasc. Pharmacol. 66:540. Towler, P. et al. (2004) J. Biol. Chem. 279:17996. 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	ACE-2, ACE-related carboxypeptidase, ACE2, ACEH, ACEHangiotensin I converting enzyme 2, Angiotensin-converting enzyme homolog, DKFZp434A014, EC 3.4.17, EC 3.4.17.23, Metalloprotease MPROT15, angiotensin I converting enzyme (peptidyl-dipeptidase A) 2, angiotensin-converting enzyme 2

Citations for Recombinant Human ACE-2 Protein, CF (24)

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Citations are publications that use Bio-Techne products. Selected citations for Recombinant Human ACE-2 Protein, CF include:

Sample Types: Recombinant Protein
Applications: Bioassay

M Haddad et al. (2022), Molecular Interactions of Tannic Acid with Proteins Associated with SARS-CoV-2 Infectivity International Journal of Molecular Sciences, 23(5).
PMID: 35269785
Species: Human
Sample Types: Protein
Applications: Bioassay

JW Kim et al. (2022), Development and Characterization of Phage-Display-Derived Novel Human Monoclonal Antibodies against the Receptor Binding Domain of SARS-CoV-2 Biomedicines, 10(12).
PMID: 36552031
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

B Bhargavan et al. (2022), SARS-CoV-2 Spike Proteins and Cell-Cell Communication Inhibits TFPI and Induces Thrombogenic Factors in Human Lung Microvascular Endothelial Cells and Neutrophils: Implications for COVID-19 Coagulopathy Pathogenesis International Journal of Molecular Sciences, 23(18).
PMID: 36142345
Species: Human
Sample Types: Protein
Applications: Bioassay

X Guo et al. (2022), Control of SARS-CoV-2 infection by MT1-MMP-mediated shedding of ACE2 Nature Communications, 13(1):7907.
PMID: 36564389
R Reindl-Sch et al. (2022), The systemic renin-angiotensin system in COVID-19 Scientific Reports, 12(1):20117.
PMID: 36418458
Species: Human
Sample Types: Protein
Applications: Bioassay

Y Polak et al. (2021), Metabolism of angiotensin peptides by angiotensin converting enzyme 2 (ACE2) and analysis of the effect of excess zinc on ACE2 enzymatic activity Peptides, 137(0):170477.
PMID: 33400951
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay

SP Shukla et al. (2021), "Molecular Masks" for ACE2 to Effectively and Safely Block SARS-CoV-2 Virus Entry International Journal of Molecular Sciences, 22(16).
PMID: 34445669
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay

T Yamaguchi et al. (2021), ACE2-like carboxypeptidase B38-CAP protects from SARS-CoV-2-induced lung injury Nature Communications, 12(1):6791.
PMID: 34815389
Applications: Enzyme Assay

J Lu et al. (2020), High affinity binding of SARS-CoV-2 spike protein enhances ACE2 carboxypeptidase activity bioRxiv, 0(0).
PMID: 32637947
Species: Canine
Sample Types: Plasma
Applications: Bioassay

É Larouche-L et al. (2020), Effect of angiotensin receptor blockers and angiotensin converting enzyme?2 on plasma equilibrium angiotensin peptide concentrations in dogs with heart disease Journal of veterinary internal medicine, 0(0).
PMID: 33368659
Species: Feline
Sample Types: Plasma
Applications: Cell Culture

T Huh et al. (2020), Effect of angiotensin receptor blockers and angiotensin-converting enzyme 2 on plasma equilibrium angiotensin peptide concentrations in cats with heart disease J Vet Intern Med, 0(0).
PMID: 33135833
Species: Human
Sample Types: Protein
Applications: Reference Standard

J Song et al. (2020), Physiological cyclic stretch up-regulates angiotensin-converting enzyme 2 expression to reduce proliferation and migration of vascular smooth muscle cells Biosci. Rep., 40(6).
PMID: 32463098
Species: Canine
Sample Types: Plasma
Applications: Bioassay

É Larouche-L et al. (2019), Plasma and tissue angiotensin-converting enzyme 2 activity and plasma equilibrium concentrations of angiotensin peptides in dogs with heart disease J. Vet. Intern. Med., 0(0).
PMID: 31254308
Species: Canine
Sample Types: Plasma
Applications: Bioassay

É Larouche-L et al. (2019), Plasma and tissue angiotensin-converting enzyme 2 activity and plasma equilibrium concentrations of angiotensin peptides in dogs with heart disease J. Vet. Intern. Med., 0(0).
PMID: 31254308
Species: Human
Sample Types: Urine
Applications: Bioassay

AM South et al. (2018), Obesity is Associated with Higher Blood Pressure and Higher Levels of Angiotensin II but Lower Angiotensin-(1-7) in Adolescents Born Preterm J. Pediatr., 0(0).
PMID: 30404738
Species: Human
Sample Types: Fluorogenic Peptide Substrate
Applications: Enzyme Assay

(2016), Angiotensin-converting enzyme 2 is reduced in Alzheimer's disease in association with increasing amyloid-? and tau pathology Alzheimers Res Ther, 8(1):50.
PMID: 27884212
Species: Human
Sample Types: Protein
Applications: Bioassay

Heurich A et al. (2014), TMPRSS2 and ADAM17 cleave ACE2 differentially and only proteolysis by TMPRSS2 augments entry driven by the severe acute respiratory syndrome coronavirus spike protein. J Virol, 88(2):1293-307.
PMID: 24227843
Species: Virus
Sample Types: Virus
Applications: Binding Assay

Zhou Y et al. (2011), A single asparagine-linked glycosylation site of the severe acute respiratory syndrome coronavirus spike glycoprotein facilitates inhibition by mannose-binding lectin through multiple mechanisms. J. Virol., 84(17):8753-64.
PMID: 20573835
Species: N/A
Sample Types: Fluorogenic Peptide Substrate
Applications: Enzyme Assay

Epelman S et al. (2008), Detection of soluble angiotensin-converting enzyme 2 in heart failure: insights into the endogenous counter-regulatory pathway of the renin-angiotensin-aldosterone system. J. Am. Coll. Cardiol., 52(9):750-4.
PMID: 18718423
Species: Primate - Chlorocebus pygerythrus (Vervet Monkey)
Sample Types: Whole Cells
Applications: Bioassay

de Lang A et al. (2006), Interferon-gamma and interleukin-4 downregulate expression of the SARS coronavirus receptor ACE2 in Vero E6 cells. Virology, 353(2):474-81.
PMID: 16860835
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry

Follis KE et al. (2006), Furin cleavage of the SARS coronavirus spike glycoprotein enhances cell-cell fusion but does not affect virion entry. Virology, 350(2):358-69.
PMID: 16519916
Species: Mouse
Sample Types: In Vivo
Applications: In Vivo

Imai Y et al. (2005), Angiotensin-converting enzyme 2 protects from severe acute lung failure. Nature, 436(7047):112-6.
PMID: 16001071
Species: Human
Sample Types: Whole Cells

Tai W et al. (0), Characterization of the receptor-binding domain (RBD) of 2019 novel coronavirus: implication for development of RBD protein as a viral attachment inhibitor and vaccine. Cell Mol Immuno17(6):613-620.
PMID: 32203189
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay

Lu J et al. (0), High affinity binding of SARS-CoV-2 spike protein enhances ACE2 carboxypeptidase activity. J Biol Che295(52):18579-18588.
PMID: 33122196

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Recombinant Human ACE-2 Protein, CF

Name: Anonymous

Application: Enzymatic activity in vitro

Reason for Rating: Very consistent results for enzymatic assay using either the fluorescent substrate Mca-APK(Dnp) or Angiotensin II.

Verified Customer | Posted 08/24/2021
Recombinant Human ACE-2 Protein, CF

Name: Anonymous

Application: In vitro bioactivity in cell culture

Verified Customer | Posted 11/21/2020
Recombinant Human ACE-2 Protein, CF

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Application: Enzymatic activity in vitro

Verified Customer | Posted 10/23/2020
Recombinant Human ACE-2 Protein, CF

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Recombinant Human ACE-2 Protein, CF

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Product Specifications for Recombinant Human ACE-2 Protein, CF

Scientific Data Examples for Recombinant Human ACE-2 Protein, CF

Formulation, Preparation and Storage

Carrier Free

Reconstitution Calculator

Background: ACE-2

Citations for Recombinant Human ACE-2 Protein, CF (24)

Customer Reviews for Recombinant Human ACE-2 Protein, CF (5)

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Recombinant Human ACE-2 Protein, CF

Recombinant Human ACE-2 Protein, CF

Recombinant Human ACE-2 Protein, CF

Recombinant Human ACE-2 Protein, CF

Recombinant Human ACE-2 Protein, CF

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