ELISA Kit for Angiotensin II (AngII)

Ang-II; Angiotensin-2

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 86-102 90
EDTA plasma(n=5) 83-97 91
heparin plasma(n=5) 79-96 78

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Angiotensin II (AngII) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Angiotensin II (AngII) 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 II (AngII) 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) 82-104% 97-105% 88-104% 94-101%
EDTA plasma(n=5) 96-105% 90-97% 91-99% 96-103%
heparin plasma(n=5) 92-102% 79-92% 97-104% 93-101%

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 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 Reference
Molecular 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
The European Journal of Clinical Investigation Mesangial medium with IgA1 from IgA nephropathy inhibits nephrin expression in mouse podocytes Wiley: source
Immunopharmacology and Inflammation Evidence for the complementary and synergistic effects of the three-alkaloid combination regimen containing berberine, hypaconitine and skimmianine on the ulcerative colitis rats induced by trinitrobenzene-sulfonic acid ScienceDirect: S0188440910001049
Nephrology Effect of aggregated immunoglobulin A1 from immunoglobulin A nephropathy patients on nephrin expression in podocytes Wiley: source
PLoS One Angiotensin II and Angiotensin-(1-7) in Paraventricular Nucleus Modulate Cardiac Sympathetic Afferent Reflex in Renovascular Hypertensive Rats PubMed: PMC3527547
BioMed Research International Heterogeneous Downregulation of Angiotensin II AT1-A and AT1-B Receptors in Arterioles in STZ-Induced Diabetic Rat Kidneys Hindawi: 947506
European Heart Journal Biglcan is beneficial in angiotensin II induced heart failure by preventing cardiac inflammation and remodeling improving LV function and mortality by preventing transdifferentiation of myofibroblasts Oxfordjournals: Source
Pflügers Archiv-European Journal of Physiology SOD1 gene transfer into paraventricular nucleus attenuates hypertension and sympathetic activity in spontaneously hypertensive rats Pubmed: 23114721
Eur J Pharmacol. Angiotensin II and vasopressin are involved in the defense system against anaphylactic hypotension in anesthetized rats Pubmed:24650734
J Mol Cell Cardiol. Increased LDL electronegativity in chronic kidney disease disrupts calcium homeostasis resulting in cardiac dysfunction Pubmed:25871829
Mol Med Rep. High glucose induces activation of the local renin?angiotensin system in glomerular endothelial cells Pubmed:24337709
Mol Med Rep. Angiotensin II promotes the progression of human gastric cancer Pubmed:24424956
PLOS ONE Intermedin in the Paraventricular Nucleus Attenuates Cardiac Sympathetic Afferent Reflex in Chronic Heart Failure Rats Pubmed:24709972
Journal of Vascular Surgery Cathepsin G deficiency reduces periaortic calcium chloride injury-induced abdominal aortic aneurysms in mice Pubmed:25037606
Biochim Biophys Acta. Cathepsin G activity lowers plasma LDL and reduces atherosclerosis Pubmed:25092171
Acta Physiologica Superoxide anions in the paraventricular nucleus mediate cardiac sympathetic afferent reflex in insulin resistance rats Pubmed:25307720
Neuroscience Letters Administration of exogenous 1,25(OH)2D3 normalizes overactivation of the central renin-angiotensin system in 1α(OH)ase knockout mice Pubmed:25576706
european heart journal A background Ca2+ PubMed: 26069213
Life Sci Cardioprotective effects of baicalein on heart failure via modulation of Ca2 PubMed: 26706290
Physiological Research Renal Sympathetic Denervation Improves Cardiac Dysfunction PubMed: 25804096
Nutrition Effects of Rubus occidentalis extract on blood pressure in subjects with prehypertension: randomized, double-blinded, placebo controlled clinical trial PubMed: 26740254
J Am Soc Hypertens Angiotensin-(1–7) enhances the effects of angiotensin II on the cardiac sympathetic afferent reflex and sympathetic activity in rostral ventrolateral medulla in renovascular hypertensive rats PubMed: 26428223
PLoS One. Up ReCavia (Guinea pig )lation of cystathione γ lyase and Hydrogen Sulphide in the Myocardium Inhibits the Progression of Isoproterenol–Caffeine Induced Left Ventricular Hypertrophy in Wistar Kyoto Rats Pubmed:26963622
Nutrition Effects of Rubus occidentalis extract on blood pressure in patients with prehypertension: Randomized, double-blinded, placebo-controlled clinical trial Pubmed:26740254
Journal of Atherosclerosis and Thrombosis Involvement of Angiotensin II Type 1 and 2 Receptors in Gelatinase ReCavia (Guinea pig )lation in Human Carotid Atheroma in vitro Pubmed:26947595
Mol Neurobiol A Therapeutic Insight of Niacin and Coenzyme Q10 Against Diabetic Encephalopathy in Rats Pubmed:26867655
Life Sci Cardioprotective effects of baicalein on heart failure via modulation of Ca(2+) handling proteins in vivo and in vitro Pubmed:26706290
PLoS One. Paradoxical Sleep Deprivation Causes Cardiac Dysfunction and the Impairment Is Attenuated by Resistance Training. pubmed:27880816
Oxidative Medicine and Cellular Longevity Epigallocatechin-3-gallate Attenuates Renal Damage by Suppressing Oxidative Stress in Diabetic db/db Mice pubmed:27698952
Journal of the Renin-Angiotensin-Aldosterone System Angiotensin type 2 receptor null mice express reduced levels of renal angiotensin converting enzyme-2/angiotensin (1-7)/Mas receptor and exhibit greater high-fat diet-induced kidney injury 1470320316661871.abstract
Microvascular Research Pleiotropic effects of survivin in vascular endothelial cells. pubmed:27373518
PLOS ONE Reduced Expression of the Extracellular Calcium-Sensing Receptor (CaSR) Is Associated withActivation of the Renin-Angiotensin System (RAS) to Promote Vascular Remodeling in the Pathogenesis of Essential Hypertension. pubmed:27391973
journal of pharmacological sciences β-Aminoisobutyric acid ameliorates the renal fibrosis in mouse obstructed kidneys via inhibitionof renal fibroblast activation and fibrosis. pubmed:28433566
Experimental Animals Ginsenoside Rb1 improves cardiac function and remodeling in heart failure. pubmed:28367863
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