ELISA Kit for Atrial Natriuretic Peptide (ANP)

ANH; ANF; CDD; Atrial Natriuretic Factor; Atrial Natriuretic Hormone; Atriopeptin; Cardionatrine; Cardiodilatin

Specificity

This assay has high sensitivity and excellent specificity for detection of Atrial Natriuretic Peptide (ANP).
No significant cross-reactivity or interference between Atrial Natriuretic Peptide (ANP) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Atrial Natriuretic Peptide (ANP) and the recovery rates were calculated by comparing the measured value to the expected amount of Atrial Natriuretic Peptide (ANP) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 84-96 89
EDTA plasma(n=5) 98-105 101
heparin plasma(n=5) 85-99 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Atrial Natriuretic Peptide (ANP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Atrial Natriuretic Peptide (ANP) 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 Atrial Natriuretic Peptide (ANP) 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) 78-99% 88-96% 89-97% 83-97%
EDTA plasma(n=5) 78-98% 91-101% 96-105% 78-95%
heparin plasma(n=5) 81-105% 97-104% 85-96% 79-95%

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
8 Volume Status and Diuretic Therapy in Systolic Heart Failure and the Detection of Early Abnormalities in Renal and Tubular Function PubMed: 21616283
Basic Research in Cardiology Release of atrial natriuretic peptide precedes shedding of the endothelial glycocalyx equally in patients undergoing on- and off-pump coronary artery bypass surgery PubMed: 21769675
Neuropsychobiology. Atrial natriuretic Peptide, arginine vasopressin Peptide and cortisol serum levels in opiate-dependent patients PubMed: 23406607
European addiction research Alcohol-induced changes in methylation status of individual CpG sites, and serum levels of vasopressin and atrial natriuretic peptide in alcohol-dependent patients during detoxification treatment. Pubmed: 24356727
Crit Care. Hypervolemia increases release of atrial natriuretic peptide and shedding of the endothelial glycocalyx Pubmed:25497357
Diabet Med.  Sodium-glucose co-transporter-2 inhibitors suppress atrial natriuretic peptide secretion in patients with newly diagnosed Type 2 diabetes. Pubmed:26927445
J Ethnopharmacol Danhong injection attenuates isoproterenol-induced cardiac hypertrophy by reCavia (Guinea pig )lating p38 and NF-κb pathway Pubmed:26970569
Journal of Babylon University/Pure and Applied Sciences Blood Pressure and Atrial Natriuretic Peptide (ANP) Levels in Patients with Autosomal Dominant Polycystic Kidney Disease. papers:uobj_paper_2016_62613672.pdf
PLoS One Enhanced heart failure, mortality and renin activation in female mice with experimental dilated cardiomyopathy pubmed:29240788
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The Journal of Biological Chemistry Toll-like receptor 2 deficiency hyperactivates the FoxO1 transcription factor and induces aging-associated cardiac dysfunction in mice Pubmed:29929978
Pediatric Cardiology Determinants of Increased Aortic Diameters in Young Normotensive Patients With Turner Syndrome Without Structural Heart Disease Pubmed:29392348
FASEB Journal Signature-oriented investigation of the efficacy of multicomponent drugs against heart failure Pubmed: 30230922
Endokrynologia Polska Endothelin-1 (ET-1), N-terminal fragment of proatrial natriuretic peptide (NTpro-ANP) and tumor necrosis factor alpha (TNF-α) in children with primary hypertension … Pubmed: 30359461
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