Wide-range ELISA Kit for Brain Natriuretic Peptide (BNP)

GC-B; B-Type Natriuretic Peptide; Ventricular Natriuretic Peptide; Gamma-brain natriuretic peptide; Brain natriuretic peptide 32

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 89-105 97
EDTA plasma(n=5) 98-105 101
heparin plasma(n=5) 80-91 82

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Wide-range Brain Natriuretic Peptide (BNP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Wide-range Brain Natriuretic Peptide (BNP) 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 Wide-range Brain Natriuretic Peptide (BNP) 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) 87-96% 78-99% 80-104% 79-98%
EDTA plasma(n=5) 85-103% 88-103% 87-97% 78-104%
heparin plasma(n=5) 83-97% 78-90% 84-95% 90-98%

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
Heart and Circulatory Physiology B-type natriuretic peptide, vascular endothelial growth factor, endothelin-1, and nitric oxide synthase in chronic mountain sickness APS: source
PLoS One Renalase deficiency in heart failure model of rats_a potential mechanism underlying circulating norepinephrine accumulation. PubMed: 21297953
International Journal of Toxicology Cardiac Troponin T and NT-proBNP as Biomarkers of Early Myocardial Damage in Amitriptyline-Induced Cardiovascular Toxicity in Rats Pubmed: 24055824
JACC ELEVATED ST2 SERUM LEVELS A BIOMARKER FOR CARDIOMYOPATHY IN DUCHENNE MUSCULAR DYSTROPHY Onlinejacc:Source
J Urol.? Why does Adenotonsillectomy Not Correct Enuresis in All Children with Sleep Disordered Breathing? Pubmed:24679871
Life Sci Cardioprotective effects of baicalein on heart failure via modulation of Ca2 PubMed: 26706290
Vlaams Diergeneeskundig Tijdschrift Equine BNP measurement using a porcine BNP enzyme-linked immunoassay: a pilot study Publication: 5950761
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Life Sci Cardioprotective effects of baicalein on heart failure via modulation of Ca(2+) handling proteins in vivo and in vitro Pubmed:26706290
J Ethnopharmacol Danhong injection attenuates isoproterenol-induced cardiac hypertrophy by reCavia (Guinea pig )lating p38 and NF-κb pathway Pubmed:26970569
Echocardiography. Improvement in the prediction of exercise-induced elevation of left ventricular filling pressure in patients with normal left ventricular ejection fraction. pubmed:27779336
SpringerPlus The effect of long-term amiodarone administration on myocardial fibrosis and evolution of left ventricular remodeling in a porcine model of ischemic … pubmed:27652141
Heart Lung Circ. Efficacy and Safety of Renal Sympathetic Denervation on Dogs with Pressure Overload-Induced Heart Failure. pubmed:27555054
Clinica Chimica Acta  Plasma concentration of fibronectin is decreased in patients with hypertrophic cardiomyopathy pubmed:27693530
Scientific Reports Paracrine cross-talk between skeletal muscle and macrophages in exercise by PGC-1α-controlled BNP. pubmed:28091624
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