Name: Recombinant Human ACE2 Protein (18-740aa), hFc-tag (Active)
Brand: CUSABIO
SKU: CSB-MP866317HU

Product Details

Description	The recombinant human ACE2 is prepared by the expression of a target fragment corresponding to 18-740aa of the human ACE2 protein with a human Fc tag at the C-terminus in the mammalian expression system. It contains endotoxin less than 1.0 EU/ug as determined by the LAL method, and its purity is over 85% as measured by SDS-PAGE. It has high reactivity and binding-affinity with SARS-CoV-2-S1 and SARS-CoV-2-S1-RBD, which were verified in the functional ELISA and LSPR assay, respectively. As is well-known, a recombinant protein tagged with Fc is relatively more stable & productive and efficient to be purified by protein A. And this ACE2 protein carries a hFc-tag, which can bind to the Fc receptor on the surface of immune cells, exerting multiple biological effects. The monocarboxypeptidase ACE2, a homolog of ACE, is a negative regulator of the renin-angiotensin-aldosterone system (RAAS). It plays a protective role in the cardiovascular system and many organs by balancing multiple functions of ACE. And ACE2 is also the primary target for SARS-CoV-2 and facilitates viral invasion into the host cells. Binding with an amino acid transporter, ACE2 is conducive to the absorption of amino acids in the kidney and intestines.
Purity	>85% (SDS-PAGE)
Endotoxin	Less than 1.0 EU/ug as determined by LAL method.
Activity	①Measured by its binding ability in a functional ELISA. Immobilized SARS-CoV-2-S (CSB-MP3324GMY) at 2 μg/ml can bind human ACE2, the EC₅₀ of human ACE2 protein is 56.64 - 103.6 ng/ml ②Measured by its binding ability in a functional ELISA. Immobilized SARS-CoV-2-S1-RBD (CSB-YP3324GMY1) at 5 μg/ml can bind human ACE2, the EC₅₀ of human ACE2 protein is 31.80 - 44.69 ng/ml. ③Measured by its binding ability in a functional ELISA. Immobilized SARS-CoV-2-S1-RBD (CSB-MP3324GMY1b1) at 5 μg/ml can bind human ACE2, the EC₅₀ is 2.785-9.139 ng/ml. ④Measured by its binding ability in a functional ELISA. Immobilized human ACE2 at 2 μg/ml can bind SARS-CoV-2-S1-RBD (CSB-MP3324GMY1), the EC₅₀ of human ACE2 protein is 8.363-12.82 ng/ml. ⑤SARS-CoV-2 Spike protein RBD his/sumostar tag (CSB-YP3324GMY1) captured on COOH chip can bind Human ACE2 protein Fc tag (CSB-MP866317HU) with an affinity constant of 100 nM as detected by LSPR Assay. ⑥SARS-CoV-2 Spike protein RBD his/myc tag (CSB-MP3324GMY1b1) captured on COOH chip can bind Human ACE2 protein Fc tag (CSB-MP866317HU) with an affinity constant of 13.8 nM as detected by LSPR Assay.
Target Names	ACE2
Uniprot No.	Q9BYF1
Alternative Names	ACE 2; ACE related carboxypeptidase; ACE-related carboxypeptidase; ACE2; ACE2_HUMAN; ACEH; Angiotensin converting enzyme 2; Angiotensin converting enzyme homolog; Angiotensin converting enzyme like protein; Angiotensin I Converting Enzyme (peptidyl dipeptidase A) 2; Angiotensin I converting enzyme 2; Angiotensin-converting enzyme homolog; DKFZP434A014; EC 3.4.17; metalloprotease MPROT 15; Metalloprotease MPROT15; OTTHUMP00000022963; Processed angiotensin-converting enzyme 2
Molecular Characterization
Species	Homo sapiens (Human)
Source	Mammalian cell
Expression Region	18-740aa
Mol. Weight	112.5 kDa
Protein Length	Partial
Tag Info	C-terminal hFc-tagged
Form	Lyophilized powder Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.
Buffer	Lyophilized from a 0.2 μm filtered PBS, 6% Trehalose, pH 7.4
Reconstitution	We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20℃/-80℃. Our default final concentration of glycerol is 50%. Customers could use it as reference.
Troubleshooting and FAQs	Protein FAQs
Storage Condition	Store at -20°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.
Shelf Life	The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself. Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.
Lead Time	Basically, we can dispatch the products out in 3-7 working days after receiving your orders. Delivery time may differ from different purchasing way or location, please kindly consult your local distributors for specific delivery time.
Notes	Repeated freezing and thawing is not recommended. Store working aliquots at 4℃ for up to one week.
Datasheet & COA	Please contact us to get it.

Customer Reviews and Q&A

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■ Q&A

What QC test will you perform?

Quality Control:
1. Cloning Stage: Sequencing of the expression plasmid to ensure that the gene is completely correct. (Free of Charge. It is our routine detection.)
2. Protein Stage:
2.1. SDS-PAGE detection to determine purity and molecular weight (determine whether the observed value and the theoretical value are consistent). We comprehensively evaluate the purity through SDS-PAGE coomassie brilliant blue staining detection and analysis using Bandscan software to ensure that the purity is more than 85%. (Free of Charge. It is our routine detection.)
2.2. Each batch will be tested for activity before shipped. (Free of Charge. It is our routine detection.)
2.3. WB detection (It is generally WB detection of tag-antibody, if there is a corresponding specific antibody, it will be arranged) to further determine the molecular weight of the protein (determine whether the observed value and the theoretical value are consistent) and its specificity. (Free of Charge. It will be performed upon customers' requests.)
2.4. AKTA-SEC test: For customers who need it, it can be used as a molecular sieve for further purification, while determining the purity and morphology of the protein (judging the presence of multimers). (Paid Service. It will be performed upon customers' requests.)
2.5. MS Identification: For customers who need it, we can arrange for Mass Spectrometry identification a separate sample. (Paid Service. It will be performed upon customers' requests.)
2.6. MS Identification: There is also a free monthly mass spectrometry identification (mixed samples will be detected). (Free of Charge. It is our routine detection.)

When making ACE2, do you use any protein denaturing purification steps?

No, we don't use any protein denaturing purification steps. The protein is purified under mild conditions to avoid denaturation of the protein and to preserve the protein's natural state to the maximum.

I would like to know if this protein has information on tests or information that can verify that there are no differences between this recombinant protein and the protein in its physiological state.

We have not tested the similarity between this mammalian cell expressed protein and native protein.
In theory, glycosylation will not be much different, and the exposed epitopes will not be very different, but the three-dimensional structure may be not the same. Because this protein is a transmembrane protein, the hydrophobic structure of the transmembrane region may affect its three-dimensional structure, but the N-terminal and C-terminal domains will generally fold correctly, so the epitope of the antigen will not change much.
HEK293 is a human embryonic kidney cell line, so it has the same immunogenicity as the native protein.

Has this protein been verified for its activity?

Please check the image part to read the validation data of this protein. It is active protein, measured by its binding ability in a functional ELISA. Immobilized SARS-CoV-2-S (CSB-MP3324GMY) at 2 μg/mL can bind human ACE2, the EC50 of human ACE2 protein is 56.64 - 103.6 ng/mL.

Do you have ACE2 protein validated with the spike protein?

Thanks for your email. We have ACE2 protein validated with the spike protein. The code is CSB-MP866317HU (human ACE2) which has been validated with CSB-MP3324GMY (human S1). Ligand Interaction Activity: Measured by its binding ability in a functional ELISA. Immobilized SARS-CoV-2-S (CSB-MP3324GMY) at 2 μg/mL can bind human ACE2, the EC50 of SARS-CoV-2-S protein is 56.64 - 103.6 ng/mL.

What's the difference between CSB-MP866317HU and CSB-AP005671HU?

Compared with CSB-AP005671HU, CSB-MP866317HU has higher activity and lower price which is an upgraded product.

Can you remove the endotoxin?

Yes. We can provide endotoxin removal service free of charge and guarantee the endotoxin level to be less than 0.1ng/μg (1EU μg).

What is the general preservative? Which kind of preservative do you usually add?

Commonly used preservative include Proclin 300, Sodium azide, etc. We do not add any preservative to our protein.

What is the general protectant? What kind of protectant do you usually add?

Commonly used protectant include saccharides, polyols, polymers, surfactants, some proteins and amino acids etc. We usually add 8% (mass ratio by volume) of trehalose and mannitol as lyoprotectant. Trehalose can significantly prevent the alter of the protein secondary structure, the extension and aggregation of proteins during freeze-drying process; mannitol is also a universal applied protectant and fillers, which can reduce the aggregation of certain proteins after lyophilization.

Can you please respond to below mentioned queries:
1) The source is Mammalian cell. It is human origin or some other mammalian cell?
2) The protein is available in lyophilized form or liquid?
3) If it is in lyophilized form, can you please provide reconstitution instructions?

1. It is human origin. We use HEK293 cell.
2. The protein is available in lyophilized form.
3. The reconstitution instruction is as below.
We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20℃/-80℃. Our default final concentration of glycerol is 50%. Customers could use it as reference.

Target Data

Function	Angiotensin-converting enzyme 2 (ACE2), also known as ACAH, is an essential counter-regulatory carboxypeptidase of the renin-angiotensin hormone system, which is a critical regulator of blood volume, systemic vascular resistance, and thus cardiovascular homeostasis. ACE2 is a suppressor of renin-angiotensin system (RAS). ACE2 converts angiotensin I to angiotensin 1-9, a nine-amino acid peptide with anti-hypertrophic effects in cardiomyocytes, and angiotensin II to angiotensin 1-7, a vasodilator. Besides the role in the cardiovascular system, ACE2 also plays a role in lung diseases as a receptor for SARS-CoV-2. Recently, with the COVID19 caused by SARS-CoV-2 spreads around the world, ACE2 functions as a receptor on the cell surface to bind with spike protein of SARS-CoV-2 in SARS-CoV-2 infection. And the mechanism of SARS-CoV-2 invasion via ACE2 are gradually being revealed. Based on these studies, there are more coronavirus-host interactome targets which have been found, such as TMPRSS2 and CD147. All of these works are critical to SARS-CoV-2 treatment, and provide potential targets of drug development for SARS-CoV-2 therapy.
Gene References into Functions	Report no influence of ACE2 gene polymorphism on stroke recurrence and only find possible interaction between hypertension history and the ACE2 gene in male stroke patients. PMID: 30056001 In a Chinese Han population, five single-nucleotide polymorphisms (SNP) (rs1514283, rs4646155, rs4646176, rs2285666, and rs879922) in ACE2 gene were determined to significantly associate with essential hypertension in female participants, while no SNP locus was linked to male group. PMID: 30335025 ACE2 and other enzymes can form ANG-(1-7) directly or indirectly from either the decapeptide ANG I or from ANG II. [review] PMID: 29351514 Suggest a potential role for ACE2 polymorphism in postexercise hypotension in hypertensive medicated individuals. PMID: 27925380 ACE2 SNPs and haplotypes are associated with circulating angiotensin-(1-7) levels. PMID: 28895159 we found an interaction between ACE2 and AGTR1 in structuralatrial fibrillation patients in a Chinese Han population PMID: 29441892 results suggest that ACE2, TNNI3K and CALM3 polymorphisms are associated with increased risk of hypertrophic cardiomyopathies and dilated cardiomyopathies and may act as disease modifiers of these diseases. PMID: 28744816 Elevated plasma ACE2 is significantly associated with more advanced LA structural remodeling in atrial fibrillation. PMID: 27738071 Overexpression of ACE2 in monocytes led to reduced endothelial adhesion, transmigration and downregulation of adhesion-related molecules and results in atherosclerosis. PMID: 28186543 These results indicated that aberrant methylation of the ACE2 promoter may be associated with EH risk. In addition, sex may significantly influence ACE2 methylation. PMID: 28440441 ACE2 rs2106809 is an important predictive factor of the response to antihypertensive treatment with ACE inhibitors in Chinese female hypertensive patients. PMID: 27121444 In islets from db/db mice, ACE2 over-expression increased intracellular calcium influx and restored impaired mitochondrial oxidation, potentially causing an increase in GSIS. These results shed light on the potential roles of ACE2 in mitochondrial metabolism, moreover, may improve our understanding of diabetes. PMID: 29128354 ACE2 may have a role in silent atherosclerosis in patients with chronic kidney disease; it counterbalances the vasoconstrictor adverse effects of angiotensin II by its conversion PMID: 27615597 This study reveals an elevated serum concentration of ACE2 and independent associations between serum ACE2 and echocardiographic parameters in hypertensive patients. PMID: 28223093 The findings of this study indicate that ACE-2 activity is reduced in AD and is an important regulator of the central classical ACE-1/Ang II/AT1R axis of renin-angiotensin system, and also that dysregulation of this pathway likely plays a significant role in the pathogenesis of Alzheimer's disease. PMID: 27884212 Overexpression of ACE2 ameliorates Abeta-induced inflammatory response by activating the ACE2/Ang-(1-7)/Mas axis in human RPE cells. PMID: 28605813 Although further studies are required to determine how clusterin suppresses non-specific cellular uptake in phagocytes, our data suggest that clusterin plays a key role in the stealth effect of not only pegylated nanoparticles but also non-pegylated nanoparticles. PMID: 27983983 The ACE2 G8790A polymorphism in type 2 diabetes mellitus patients was correlated with cerebral stroke, and the A allele might be a risk factor of type 2 diabetes mellitus combined with cerebral stroke. PMID: 27500554 ollectrin, an ACE2 homolog with no catalytic activity, regulates blood pressure through an NO-dependent mechanism. Large body of experimental data confirmed sustained beneficial effects of ACE2/Ang-(1-7)/Mas receptor axis activation on hypertension and vascular injury. PMID: 27889958 The potential contribution of the ACE2 to cardiovascular disease progression was addressed. PMID: 27965422 Serum ACE2 activity was significantly lower in acute ischemic stroke as compared to both control and stroke-alert patients, followed by an increase to control levels at three days. PMID: 27488276 Here, we review the role and effects of ACE2, ACE2 activators, Ang-(1-7) and synthetic Mas receptor agonists in the control of inflammation and fibrosis in cardiovascular and renal diseases and as counter-regulators of the ACE-Ang II-AT1 axis. PMID: 26995300 ACE2 overexpression inhibited cell growth. PMID: 27460845 Downregulation of ACE2/Ang-(1-7)/Mas axis stimulates breast cancer metastasis through the activation of store-operated calcium entry and PAK1/NF-kappaB/Snail1 pathways. PMID: 27063099 The circulating ACE2 and Ang-(1-7) levels were related to neither rs4646155 nor rs879922 in female or male patients.In conclusion, the rs2106809 polymorphism of the ACE2 gene may be a determinant of the circulating Ang-(1-7) level in female patients with hypertension, suggesting a genetic association between circulating Ang-(1-7) levels and ACE2 gene polymorphisms in patients with hypertension. PMID: 27310975 These results indicated that angiotensin-(1-7)/ACE2/Mas axis may reduce liver lipid accumulation partly by regulating lipid-metabolizing genes through ATP/P2 receptor/CaM signaling pathway. PMID: 26883384 ACE-2 significantly increased when IMR-90 were hypoxic prior to hyperoxic exposure with no recovery. PMID: 27093376 Imbalanced down-regulation of ACE and ACE2 mRNA expression levels may play an important role in the development and progression of thoracic aortic aneurysmal dilatation and subsequently dissection. PMID: 25237166 These results suggest that the ACE2 G8790A and rs2106809 polymorphisms may be associated with essential hypertension risk. PMID: 25237167 ACE and ACE2 expression at the mRNA and protein levels are significantly increased in the myocardium of patients with heart failure. PMID: 25869724 Multivariable regression analysis revealed that urinary L-FABP and urinary albumin/ creatinine ratio were significantly associated with urinary ACE2 levels. PMID: 26067610 ACE2 and Ang-(1-7) significantly inhibit early atherosclerotic lesion formation via protection of endothelial function and inhibition of inflammatory response. PMID: 25721616 ACE-2 is expressed in fetal human lung fibroblasts but is significantly decreased by hyperoxic gas PMID: 25665060 urinary ACE2 increased in type 2 diabetic patients with various degrees of albuminuria PMID: 25791940 soluble ACE2 is involved in the pathomechanism of hypertension and heart failure. PMID: 24691269 Decrease in circulating ACE2 activity was associated with cardiovascular disease in patients with chronic kidney disease. PMID: 25813276 By genetic replenishment of ACE2 and pharmaceutical use of ARB, restored ACE2 level mitigated GBC growth. Our results supported the rationale for the use of ARB in GBC patients for potential therapeutic benefit PMID: 25663464 There is no clear association between ACE2 gene A9570G polymorphisms and childhood primary nephrotic syndrome. PMID: 25815490 Our results revealed that SNPs rs2074192 and rs714205 in ACE2 gene were associated with the susceptibility of DR and PDR. PMID: 25359286 Olmesartan may uniquely increase urinary ACE2 level in hypertensive patients, which could potentially offer additional renoprotective effects. PMID: 24842388 Hepatocellular carcinoma patients with higher level of ACE2 expression had longer survival time than those with lower level of ACE2 expression. PMID: 25701390 Urinary ACE2 activity and protein expression are increased in type 1 diabetes patients prior to the onset of clinical complications. PMID: 24920267 Brain endoplasmic reticulum stress does not contribute to DOCA-salt hypertension and ACE2 blunts neurogenic hypertension independently of ER stress. PMID: 25519733 Partial loss of ACE2 is sufficient to enhance the susceptibility to heart disease. PMID: 24728465 These data demonstrate that MSCs modified to overexpress the ACE2 gene can produce biologically active ACE2 protein over a sustained period of time and have an enhanced ability to promote endothelial repair after LPS challenge PMID: 25200929 ACE2 cleavage is regulated by influenza A H1N1 neuraminidase. PMID: 24662240 among females ACE2 rs2106809 polymorphisms,and among males ACE2 rs2106809 polymorphism and alcohol consumption are associated with essential hypertension PMID: 24112034 Data suggest angiotensin (ANG) converting enzyme 2/ANG-II-(1-7)/proto-oncogene protein Mas axis regulates inflammation/fibrosis, cell proliferation, and leukocyte recruitment/activation; main topic here is kidney/inflammatory renal disease. [REVIEW] PMID: 23488800 ACE2 and FZD1 are prognosis markers in squamous cell/adenosquamous carcinoma and adenocarcinoma of gallbladder. PMID: 23921915 ACE2 overexpression in the rostral ventrolateral medulla attenuates the enhanced tonically active glutamatergic input in SHRs, which may be an important mechanism underlying the beneficial effect of central ACE2 to hypertension. PMID: 24838502 Show More Hide All
Subcellular Location	Processed angiotensin-converting enzyme 2: Secreted, SUBCELLULAR LOCATION: Cell membrane, Single-pass type I membrane protein, Cytoplasm
Protein Families	Peptidase M2 family
Tissue Specificity	Expressed in endothelial cells from small and large arteries, and in arterial smooth muscle cells. Expressed in lung alveolar epithelial cells, enterocytes of the small intestine, Leydig cells and Sertoli cells (at protein level). Expressed in heart, kidn
Database Links	HGNC: 13557 OMIM: 300335 KEGG: hsa:59272 STRING: 9606.ENSP00000252519 UniGene: Hs.178098

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Code	CSB-MP866317HU
Size
Image	The recombinant human ACE2 (Fc Tag) protein has a calculated molecular weight of 112.5 kDa. Due to glycosylation, the rhACE2/Fc migrates on SDS-PAGE under reducing conditions, forming a protein band of about 110-120 kDa. Activity Assay- ELISA Measured by ACE2 protein (CSB-MP866317HU) binding ability in a functional ELISA. Coat the plate with 0.2μg/well (2μg/ml, 100μl/well) of SARS-CoV-2 S1 protein (CSB-MP3324GMY), add 100μl/well (0.7629-100000ng/ml, gradient dilution, 2x to 18x) of ACE2 protein and 100μl/well of Peroxidase-Affinipure Goat Anti-Human IgG, Fcγ Fragment Specific (CSB-PA160506). Wells were washed with buffer, and then A450 nm was measured. The results showed SARS-CoV-2 S1 protein could bind ACE2 protein. The EC50 is 56.64 - 103.6 ng/ml. Biological Activity Assay Activity Assay- ELISA Measured by ACE2 protein (CSB-MP866317HU) binding ability in a functional ELISA. Coat the plate with 0.5μg/well (5μg/ml, 100μl/well) of SARS-CoV-2 S1-RBD protein (CSB-YP3324GMY1), add 100μl/well (0.762939-100000ng/ml, gradient dilution, 2x to 18x) of ACE2 protein and 100μl/well of Peroxidase-Affinipure Goat Anti-Human IgG, Fcγ Fragment Specific (CSB-PA160506). Wells were washed with buffer, and then A450 nm was measured. The results showed SARS-CoV-2 S1-RBD protein could bind ACE2 protein. The EC50 is 31.80 - 44.69 ng/ml. Biological Activity Assay Activity Assay- ELISA Measured by ACE2 protein (CSB-MP866317HU) binding ability in a functional ELISA. Coat the plate with 0.5μg/well (5μg/ml, 100μl/well) of SARS-CoV-2 S1-RBD protein (CSB-MP3324GMY1b1), add 100μl/well (0.762939-100000ng/ml, gradient dilution, 2x to 18x) of ACE2 protein and 100μl/well of Peroxidase-Affinipure Goat Anti-Human IgG, Fcγ Fragment Specific (CSB-PA160506). Wells were washed with buffer, and then A450 nm was measured. The results showed SARS-CoV-2 S1-RBD protein could bind ACE2 protein. The EC50 is 2.785-9.139 ng/ml. Biological Activity Assay Activity Assay- ELISA Measured by ACE2 protein (CSB-MP866317HU) binding ability in a functional ELISA. Coat the plate with 0.2μg/well (2μg/ml, 100μl/well) of ACE2 protein, add 100μl/well (0.00001164153-200000ng/ml, gradient dilution, 4x to 18x) of SARS-CoV-2 S1-RBD protein (CSB-MP3324GMY1) and 100μl/well of Peroxidase-Affinipure Goat Anti-Human IgG, Fcγ Fragment Specific (CSB-PA160506). Wells were washed with buffer, and then A450 nm was measured. The results showed SARS-CoV-2 S1-RBD protein could bind ACE2 protein. The EC50 is 8.363-12.82 ng/ml ng/ml. Biological Activity Assay LSPR Assay The SPR sensorgram shows the binding between ACE2 protein (CSB-MP866317HU) and SARS-CoV-2 S1-RBD protein (CSB-YP3324GMY1). ACE2 protein was injected at 5 concentrations (400, 200, 100, 50, 0 nM) across 0.5 mg/ml of SARS-CoV-2 S1-RBD protein captured on a COOH chip. The results showed ACE2 protein could bind SARS-CoV-2 S1-RBD protein with an affinity constant of 100 nM. Biological Activity Assay LSPR Assay The SPR sensorgram shows the binding between ACE2 protein (CSB-MP866317HU) and SARS-CoV-2 S1-RBD protein (CSB-MP3324GMY1b1). ACE2 protein was injected at 5 concentrations (100, 50, 25, 12.5, 0 nM) across 0.5 mg/ml of SARS-CoV-2 S1-RBD protein captured on a COOH chip. The results showed ACE2 protein could bind SARS-CoV-2 S1-RBD protein with an affinity constant of 13.8 nM. Biological Activity Assay
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Recombinant Human ACE2 Protein (18-740aa), hFc-tag (Active)

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