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.

MatrixRecovery range (%)Average(%)
serum(n=5)86-10290
EDTA plasma(n=5)83-9791
heparin plasma(n=5)79-9678

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.

Sample1:21:41:81:16
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

ReagentsQuantityReagentsQuantity
Pre-coated, ready to use 96-well strip plate1Plate sealer for 96 wells4
Standard2Standard Diluent1×20mL
Detection Reagent A1×120µLAssay Diluent A1×12mL
Detection Reagent B1×120µLAssay Diluent B1×12mL
TMB Substrate1×9mLStop Solution1×6mL
Wash Buffer (30 × concentrate)1×20mLInstruction manual1

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.

Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Angiotensin II (AngII) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Angiotensin II (AngII) and unlabeled Angiotensin II (AngII) (Standards or samples) with the pre-coated antibody specific to Angiotensin II (AngII). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Angiotensin II (AngII) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Angiotensin II (AngII) in the sample.

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INCREMENT SERVICES

MagazineReference
Molecular medicine Angiotensin-Converting Enzyme 2 Overexpression Remarkably Ameliorated Glomerular Injury in a Rat Model of Diabetic Nephropathy: A Comparison with ACE InhibitionMolMed: 10_11_liu
PLoS OneAngiotensin II and Angiotensin-(1-7) in Paraventricular Nucleus Modulate Cardiac Sympathetic Afferent Reflex in Renovascular Hypertensive RatsPubMed: PMC3527547
BioMed Research InternationalHeterogeneous Downregulation of Angiotensin II AT1-A and AT1-B Receptors in Arterioles in STZ-Induced Diabetic Rat KidneysHindawi: 947506
Pflügers Archiv-European Journal of PhysiologySOD1 gene transfer into paraventricular nucleus attenuates hypertension and sympathetic activity in spontaneously hypertensive ratsPubmed: 23114721
Eur J Pharmacol. Angiotensin II and vasopressin are involved in the defense system against anaphylactic hypotension in anesthetized ratsPubmed:24650734
Mol Med Rep.High glucose induces activation of the local renin?angiotensin system in glomerular endothelial cellsPubmed:24337709
PLOS ONEIntermedin in the Paraventricular Nucleus Attenuates Cardiac Sympathetic Afferent Reflex in Chronic Heart Failure RatsPubmed:24709972
Acta PhysiologicaSuperoxide anions in the paraventricular nucleus mediate cardiac sympathetic afferent reflex in insulin resistance ratsPubmed:25307720
Life SciCardioprotective effects of baicalein on heart failure via modulation of Ca2PubMed: 26706290
Physiological ResearchRenal Sympathetic Denervation Improves Cardiac DysfunctionPubMed: 25804096
J Am Soc HypertensAngiotensin-(1–7) enhances the effects of angiotensin II on the cardiac sympathetic afferent reflex and sympathetic activity in rostral ventrolateral medulla in renovascular hypertensive ratsPubMed: 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 RatsPubmed:26963622
Mol NeurobiolA Therapeutic Insight of Niacin and Coenzyme Q10 Against Diabetic Encephalopathy in RatsPubmed:26867655
Life SciCardioprotective effects of baicalein on heart failure via modulation of Ca(2+) handling proteins in vivo and in vitroPubmed:26706290
PLoS One. Paradoxical Sleep Deprivation Causes Cardiac Dysfunction and the Impairment Is Attenuated by Resistance Training.pubmed:27880816
Microvascular ResearchPleiotropic effects of survivin in vascular endothelial cells.pubmed:27373518
PLOS ONEReduced 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 sciencesβ-Aminoisobutyric acid ameliorates the renal fibrosis in mouse obstructed kidneys via inhibitionof renal fibroblast activation and fibrosis.pubmed:28433566
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