ELISA Kit for Angiotensin 1-7 (Ang1-7)

Specificity

This assay has high sensitivity and excellent specificity for detection of Angiotensin 1-7 (Ang1-7).
No significant cross-reactivity or interference between Angiotensin 1-7 (Ang1-7) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Angiotensin 1-7 (Ang1-7) and the recovery rates were calculated by comparing the measured value to the expected amount of Angiotensin 1-7 (Ang1-7) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 94-103 97
EDTA plasma(n=5) 80-89 79
heparin plasma(n=5) 86-94 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Angiotensin 1-7 (Ang1-7) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Angiotensin 1-7 (Ang1-7) 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 1-7 (Ang1-7) 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) 93-101% 79-98% 95-105% 78-105%
EDTA plasma(n=5) 95-103% 96-104% 79-105% 96-104%
heparin plasma(n=5) 83-96% 92-104% 92-101% 89-97%

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 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
J Stroke Cerebrovasc Dis Improvement of Plasma Biomarkers after Switching Stroke Patients from Other Angiotensin II Type I Receptor Blockers to Olmesartan PubMed: 25891757
Cellular Physiology and Biochemistry Angiotensin-(1-7) in Paraventricular Nucleus Contributes to the Enhanced Cardiac Sympathetic Afferent Reflex and Sympathetic Activity in Chronic Heart Failure Rats pubmed:28848201
Frontiers in Physiology Hydrogen Sulfide Attenuates Atherosclerosis in a Partially Ligated Carotid Artery Mouse model via Regulating Angiotensin Converting Enzyme 2 Expression pubmed:29066981
Molecular Medicine Reports The ACE2-Ang (1-7)-Mas receptor axis attenuates cardiac remodeling and fibrosis in post-myocardial infarction. pubmed:28656296
Journal of Clinical Pharmacy and Therapeutics Relationship between genetic variants of ACE2 gene and circulating levels of ACE2 and its metabolites. pubmed:28895159
Molecular Medicine Reports Effects of IQP, VEP and Spirulina platensis hydrolysates on the local kidney renin angiotensin system in spontaneously hypertensive rats pubmed:28944898
Open Journal of Endocrine and Metabolic Diseases Paper Submission A Study of the Correlation between Angiotensin (1-7) and the Histopathological Changes in the Penises of Experimentally Diabetic Rats 2-1980272_83198.htm
Journal of the American Society of Hypertension Mesenchymal stem cell–derived microvesicles alleviate pulmonary arterial hypertension by regulating renin-angiotensin system Pubmed:29752040
Innate Immunity  Lipoxin A attenuates LPS-induced acute lung injury via activation of the ACE2-Ang-(1-7)-Mas axis Pubmed:29969931
Evidence-based Complementary and Alternative Medicine  Acute Biochemical, Cardiovascular, and Autonomic Response to Hyperbaric (4 atm) Exposure in Healthy Subjects Pubmed:29977313
European Journal of Obstetrics & Gynecology and Reproductive Biology The impact of ovulation inducing drugs on carcinogenesis
European Journal of Obstetrics & Gynecology and Reproductive Biology Changes of angiotensin II (1–8) and angiotensin (1–7) levels in early and late-onset preeclampsia
Journals of Gerontology Series A-Biological Sciences and Medical Sciences Therapeutic Delivery of Ang (1-7) Via Genetically Modified Probiotic: A Dosing Study. Pubmed: 31586210
journal of the european academy of dermatology and venereology Significance of the angiotensin I/angiotensin II/angiotensin‐(1‐7) axis in the pathogenesis of systemic sclerosis Pubmed: 31746507
Medicine Alteration and association between serum ACE2/angiotensin (1-7)/Mas axis and oxidative stress in chronic kidney disease: A pilot study Pubmed: 32756181
Dikkat Eksikliği Hiperaktivite Bozukluğu Tanısı Olan Anjiyotensin I, II, III, Alamandin peptid, Anjiyotensin Dönüştürücü Enzim (ACE), NADPH Oksidaz Enzim Serum …
Amino Acids Angiotensin-(1¨C7) attenuates collagen-induced arthritis via inhibiting oxidative stress in rats 33398523
Autoimmunity The prevalence and role of functional autoantibodies to angiotensin-converting-enzyme-2 in patients with systemic sclerosis 33910447
researchsquare Ang-(1-7) Protects Skeletal Muscle Function in Aged Mice
Peptides Angiotensin-(1-7) protects against sepsis-associated left ventricular dysfunction induced by lipopolysaccharide 34298021
BMC Musculoskeletal Disorders Ang-(1–7) protects skeletal muscle function in aged mice 34548056
J Cardiovasc Pharmacol Resveratrol Confers Protection Against Ischemia/Reperfusion Injury by Increase of Angiotensin (1-7) Expression in a Rat Model of Myocardial Hypertrophy 34232225
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