ELISA Kit for Angiotensin I Converting Enzyme 2 (ACE2)

ACEH; ACEII; ACE-II; Peptidyl-Dipeptidase A; ACE-related carboxypeptidase; Angiotensin-converting enzyme homolog; Metalloprotease MPROT15

Specificity

This assay has high sensitivity and excellent specificity for detection of Angiotensin I Converting Enzyme 2 (ACE2).
No significant cross-reactivity or interference between Angiotensin I Converting Enzyme 2 (ACE2) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Angiotensin I Converting Enzyme 2 (ACE2) and the recovery rates were calculated by comparing the measured value to the expected amount of Angiotensin I Converting Enzyme 2 (ACE2) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 99-105 102
EDTA plasma(n=5) 79-93 80
heparin plasma(n=5) 78-95 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Angiotensin I Converting Enzyme 2 (ACE2) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Angiotensin I Converting Enzyme 2 (ACE2) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Angiotensin I Converting Enzyme 2 (ACE2) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 84-92% 83-97% 80-95% 97-104%
EDTA plasma(n=5) 80-102% 91-101% 78-104% 83-105%
heparin plasma(n=5) 88-104% 85-92% 98-105% 78-101%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Nature Medicine Angiotensin-Converting Enzyme 2 Overexpression Remarkably Ameliorated Glomerular Injury in a Rat Model of Diabetic Nephropathy: A Comparison with ACE Inhibition MolMed: 10_11_liu
Journal of Cancer Therapy Plasma Levels of Angiotensin-Converting Enzymes 1 and 2 and AGTR2 (T1247G and A5235G) Gene Polymorphisms Are Associated to Breast Cancer Progression. Scirp: Source
Reprod Toxicol.? ACE2 activation by xanthenone prevents leptin-induced increases in blood pressure and proteinuria during pregnancy in Sprague-Dawley rats Pubmed:25205467
Intensive Care Medicine Experimental Angiotensin converting enzymes in patients with acute respiratory distress syndrome Content: 3
Journal of Chromatography A Application of a new procedure for liquid chromatography/mass spectrometry profiling of plasma amino acid-related metabolites and untargeted shotgun proteomics to identify mechanisms and biomarkers of calcific aortic stenosis S0021967317311858
Journal of Clinical Pharmacy and Therapeutics Relationship between genetic variants of ACE2 gene and circulating levels of ACE2 and its metabolites. pubmed:28895159
Innate Immunity  Lipoxin A attenuates LPS-induced acute lung injury via activation of the ACE2-Ang-(1-7)-Mas axis Pubmed:29969931
Journal of Surgical Research Impact of angiotensin-converting enzyme 2 levels on postoperative pneumonia after esophagectomy Pubmed:29506841
canadian journal of physiology and pharmacology Could cardioprotective effect of ACE2 activator “diminazene aceturate” is more potent than ACE inhibitor “Enalapril” on acute myocardial infarction in rats? Pubmed: 30840489
Intensive Care Medicine Angiotensin-converting enzymes in acute respiratory distress syndrome
BioMed Research International AVE 0991 Attenuates Pyroptosis and Liver Damage after Heatstroke by Inhibiting the ROS-NLRP3 Inflammatory Signalling Pathway
Heart and vessels Relationship between circulating levels of angiotensin-converting enzyme 2-angiotensin-(1–7)-MAS axis and coronary heart disease Pubmed: 31359146
Medicine Alteration and association between serum ACE2/angiotensin (1-7)/Mas axis and oxidative stress in chronic kidney disease: A pilot study Pubmed: 32756181
American Journal of Respiratory and Critical Care Medicine Equilibrium Angiotensin Metabolite Profiling in Patients with Acute Respiratory Distress Syndrome Indicates Angiotensin Converting Enzyme Inhibition Pubmed: 32628511
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