Human NT Pro-BNP DuoSet ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
(NT Pro-BNP). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, and plasma samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Kit Content
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Scientific Data
Product Datasheets
Preparation and Storage
Background: BNP
Brain-type Natriuretic Peptide (BNP) is a nonglycosylated peptide that is produced predominantly by ventricular myocytes and belongs to the natriuretic peptide family. Proteolytic cleavage of the 12 kDa BNP precursor gives rise to N-terminal proBNP (NT-proBNP) and mature BNP. Plasma NT-proBNP is a marker for congestive heart failure, while mature BNP (aa 103 - 134) promotes vasodilation and fluid and sodium excretion. Human BNP precursor shares 29% and 51% aa sequence identity with mouse and porcine BNP precursor, respectively.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Citations for Human NT Pro-BNP DuoSet ELISA
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 10
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Comparative Effect of Two Types of Physical Exercise for the Improvement of Exercise Capacity, Diastolic Function, Endothelial Function and Arterial Stiffness in Participants with Heart Failure with Preserved Ejection Fraction (ExIC-FEp Study): Protocol for a Randomized Controlled Trial
Authors: Cavero-Redondo, I;Saz-Lara, A;Mart�nez-Garc�a, I;Bizzozero-Peroni, B;D�az-Go�i, V;D�ez-Fern�ndez, A;Moreno-Herr�iz, N;Pascual-Morena, C;
Journal of clinical medicine
Species: Human
Sample Types: Plasma
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Serum Pro-N-Cadherin Is a Marker of Subclinical Heart Failure in the General Population
Authors: PD Ferrell, KM Oristian, I Puranam, SV Pizzo
Journal of the American Heart Association, 2023-03-09;0(0):e028234.
Species: Human
Sample Types: Serum
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Treatment of Acute Respiratory Distress Syndrome Caused by COVID-19 with Human Umbilical Cord Mesenchymal Stem Cells
Authors: T Bukreieva, H Svitina, V Nikulina, A Vega, O Chybisov, I Shablii, A Ustymenko, P Nemtinov, G Lobyntseva, I Skrypkina, V Shablii
International Journal of Molecular Sciences, 2023-02-23;24(5):.
Species: Human
Sample Types: Plasma
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Clinical and prognostic relevance of sST2 in adults with dengue-associated cardiac impairment and severe dengue
Authors: A Teo, PY Chia, GK Ramireddi, SKM Khoo, TW Yeo
PloS Neglected Tropical Diseases, 2022-10-13;16(10):e0010864.
Species: Human
Sample Types: Plasma
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Intact Fibroblast Growth Factor 23 Regulates Chronic Kidney Disease-Induced Myocardial Fibrosis by Activating the Sonic Hedgehog Signaling Pathway
Authors: L Li, H Gan
Journal of the American Heart Association, 2022-09-14;0(0):e026365.
Species: Rat
Sample Types: Serum
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Biomarkers of Pulmonary Hypertension Are Altered in Children With Down Syndrome and Pulmonary Hypertension
Authors: M Griffiths, J Yang, D Vaidya, M Nies, S Brandal, DD Ivy, F Hickey, K Wolter-War, ED Austin, M Mullen, MW Pauciulo, KA Lutz, EB Rosenzweig, R Hirsch, D Yung, WC Nichols, AD Everett
The Journal of pediatrics, 2021-10-21;0(0):.
Species: Human
Sample Types: Plasma
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Artificial Intelligence-Assisted Identification of Genetic Factors Predisposing High-Risk Individuals to Asymptomatic Heart Failure
Authors: NI Yang, CH Yeh, TH Tsai, YJ Chou, PW Hsu, CH Li, YH Chan, LT Kuo, CT Mao, YC Shyu, MJ Hung, CC Lai, HK Sytwu, TF Tsai
Cells, 2021-09-15;10(9):.
Species: Human
Sample Types: Plasma
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Alterations of Cardiac KATP Channels and Autophagy Contribute in the Late Cardioprotective Phase of Exercise Preconditioning
Authors: J Lu, SS Pan, QT Wang, Y Yuan
Int Heart J, 2018-08-11;0(0):.
Species: Rat
Sample Types: Serum
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The DiPEP (Diagnosis of PE in Pregnancy) biomarker study: An observational cohort study augmented with additional cases to determine the diagnostic utility of biomarkers for suspected venous thromboembolism during pregnancy and puerperium
Authors: BJ Hunt, K Parmar, K Horspool, N Shephard, C Nelson-Pie, S Goodacre
Br. J. Haematol., 2018-01-23;0(0):.
Species: Human
Sample Types: Serum
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Comparison of change in end tidal carbon dioxide after three minutes of step exercise between systemic sclerosis patients with and without pulmonary hypertension
Authors: Elana J Bernstein
Rheumatology (Oxford), 2016-10-25;56(1):87-94.
Species: Human
Sample Types: Serum
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We used human serum with no dilution. Good ELISA.
