Multiplex Assay Kit for Atrial Natriuretic Peptide (ANP) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

(Note: Up to 8-plex in one testing reaction)

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-101 87
EDTA plasma(n=5) 83-94 87
heparin plasma(n=5) 93-102 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Atrial Natriuretic Peptide (ANP) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 96-103% 92-101% 92-101% 90-99%
EDTA plasma(n=5) 92-103% 82-102% 79-101% 90-101%
heparin plasma(n=5) 87-102% 93-105% 78-93% 78-94%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:ANP) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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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
Drug Delivery Ginsenoside Rg3 micelles mitigate doxorubicin-induced cardiotoxicity and enhance its anticancer efficacy. pubmed:29063791
Elife SIRT2 deacetylase regulates the activity of GSK3 isoforms independent of inhibitory phosphorylation Pubmed:29504933
Science of the Total Environment Co-exposure of silica nanoparticles and methylmercury induced cardiac toxicity and Pubmed:29727991
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
International Journal of Peptide Research and Therapeutics ANP/NPRA Inhibits Epithelial-Mesenchymal Transition of Airway by Targeting Smad3 in Asthma
Van Veterinary Journal Relationship between Natriuretic Peptide Levels with Heart and Kidney Parameters in Rats with Chronic Renal Failure.
Acta Pharmaceutica Sinica B The poly (ADP-ribosyl) ation of BRD4 mediated by PARP1 promoted pathological cardiac hypertrophy 34094834
Peptides Alarin moderated myocardial hypertrophy via inhibiting cyclic adenosine monophosphate/protein kinase A signaling pathway to attenuate autophagy 34624429
Peptides Osteocrin alleviates cardiac hypertrophy via attenuating oxidative stress Pubmed:35218795
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