ELISA Kit for Atrial Natriuretic Peptide (ANP) Canis familiaris; Canine (Dog) Competition ELISA

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

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  • ELISA Kit for Atrial Natriuretic Peptide (ANP) Packages (Simulation)
  • ELISA Kit for Atrial Natriuretic Peptide (ANP) Packages (Simulation)
  • ELISA Kit for Atrial Natriuretic Peptide (ANP) Results demonstration
  • CEA225Ca.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

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) 80-88 85
EDTA plasma(n=5) 88-103 98
heparin plasma(n=5) 84-103 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) 88-96% 98-105% 88-95% 95-102%
EDTA plasma(n=5) 93-105% 78-97% 79-90% 92-104%
heparin plasma(n=5) 96-105% 93-103% 81-102% 85-94%

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.

ELISA Kit for Atrial Natriuretic Peptide (ANP)

Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Atrial Natriuretic Peptide (ANP) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Atrial Natriuretic Peptide (ANP) and unlabeled Atrial Natriuretic Peptide (ANP) (Standards or samples) with the pre-coated antibody specific to Atrial Natriuretic Peptide (ANP). 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 Atrial Natriuretic Peptide (ANP) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Atrial Natriuretic Peptide (ANP) in the sample.

Citations

  • Volume Status and Diuretic Therapy in Systolic Heart Failure and the Detection of Early Abnormalities in Renal and Tubular FunctionPubMed: 21616283
  • Release of atrial natriuretic peptide precedes shedding of the endothelial glycocalyx equally in patients undergoing on- and off-pump coronary artery bypass surgeryPubMed: 21769675
  • Atrial natriuretic Peptide, arginine vasopressin Peptide and cortisol serum levels in opiate-dependent patientsPubMed: 23406607
  • 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
  • Hypervolemia increases release of atrial natriuretic peptide and shedding of the endothelial glycocalyxPubmed:25497357
  • Sodium-glucose co-transporter-2 inhibitors suppress atrial natriuretic peptide secretion in patients with newly diagnosed Type 2 diabetes.Pubmed:26927445
  • Danhong injection attenuates isoproterenol-induced cardiac hypertrophy by reCavia (Guinea pig )lating p38 and NF-κb pathwayPubmed:26970569
  • Blood Pressure and Atrial Natriuretic Peptide (ANP) Levels in Patients with Autosomal Dominant Polycystic Kidney Disease.papers:uobj_paper_2016_62613672.pdf
  • Enhanced heart failure, mortality and renin activation in female mice with experimental dilated cardiomyopathypubmed:29240788
  • Ginsenoside Rg3 micelles mitigate doxorubicin-induced cardiotoxicity and enhance its anticancer efficacy.pubmed:29063791
  • SIRT2 deacetylase regulates the activity of GSK3 isoforms independent of inhibitory phosphorylationPubmed:29504933
  • Co-exposure of silica nanoparticles and methylmercury induced cardiac toxicity andPubmed:29727991
  • Toll-like receptor 2 deficiency hyperactivates the FoxO1 transcription factor and induces aging-associated cardiac dysfunction in micePubmed:29929978
  • Determinants of Increased Aortic Diameters in Young Normotensive Patients With Turner Syndrome Without Structural Heart DiseasePubmed:29392348
  • Signature-oriented investigation of the efficacy of multicomponent drugs against heart failurePubmed: 30230922
  • 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
  • ANP/NPRA Inhibits Epithelial-Mesenchymal Transition of Airway by Targeting Smad3 in Asthma
  • Relationship between Natriuretic Peptide Levels with Heart and Kidney Parameters in Rats with Chronic Renal Failure.
  • The poly (ADP-ribosyl) ation of BRD4 mediated by PARP1 promoted pathological cardiac hypertrophy34094834
  • Alarin moderated myocardial hypertrophy via inhibiting cyclic adenosine monophosphate/protein kinase A signaling pathway to attenuate autophagy34624429
  • Osteocrin alleviates cardiac hypertrophy via attenuating oxidative stressPubmed:35218795

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