NT-proBNP ELISA Assay Kit

$610.00

The Eagle Biosciences NT-proBNP ELISA Assay Kit (enzyme-linked immunoassay kit) is intended for the quantitative determination of human Nt-proBNP in serum. The NT-proBNP ELISA Assay Kit is manufactured by Biomedica (www.bmgrp.com).  The NT-proBNP ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

NT-proBNP ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 3 pmol/L
Dynamic Range: 0 – 640 pmol/L
Incubation Time: 3.5 hours
Sample Type: Serum
Sample Size: 50 µL
Species: Human

Controls Included


Product Developed and Manufactured in Austria by Biomedica


Learn more about BNP at Eagle Biosciences Biomarker Spotlight page dedicated to BNP here: BNP Biomarker Spotlight

Additional Information

Assay Background

The NT-proBNP ELISA Assay kit is an imunoassay for the quantitative determination of NT-proBNP in Serum. The mature form of brain natriuretic peptide (BNP) is formed from the BNP (1-108) precursor and is secreted predominantly by the cardiac ventricles. Circulating forms of BNP include BNP-32 (77-108), a high molecular weight BNP which has not been well characterizedand and the nt-proBNP (1-76). The BNP fragment is typically measured as a proxy for BNP because of its longer half-life. Recent research suggests that natriuretic peptides are important protectors against fluid overload and high blood pressure. BNP has also been implicated as a valuable marker in assessing cardiac risk.
The natriuretic peptides are members of a family of structurally similar but genetically distinct peptide hormones, consisting of atrial-, brain-, and C-type (ANP, BNP, and CNP, respectively). ANP and BNP preferentially bind to a membrane-bound guanylyl cyclase (GC) receptor called GC-A or NPR1, whereas CNP is the physiological ligand for GC-B (NPR2). The natriuretic peptides play an important role in the regulation of cardiovascular and renal homeostasis and in the regulation of fatty acid metabolism and body weight.
BNP is mainly expressed by ventricular myocardium in response to volume overload and increased filling pressure. BNP has a cleavable signal sequence. Mature BNP consists of 108 amino acids (proBNP or BNP-108), and undergoes cleavage resulting in physiologically active BNP-32 and additional C-terminal fragments along with a physiologically inactive N-terminal peptide comprising amino acids 1-76, which is further degraded proteolytically. BNP fragments in the circulation are therefore very heterogenous.
BNP has a key role in cardiovascular homeostasis with biological actions including natriuresis, diuresis, vasorelaxation, and inhibition of renin and aldosterone secretion. A high concentration of BNP in the bloodstream is indicative of heart failure. The discovery of natriuretic peptides identified an endocrine system that contributes to diuresis and vascular tone. The biology, biochemistry and the pathophysiological role of natriuretic peptides are described in several reviews.

Assay Principle

  • All reagents and samples must be at room temperature (18-26°C) before use in the NT-proBNP ELISA Assay Kit.
  • Mark position for BLANK/STD/SAMPLE/CTRL (Blank/Standard/Sample/Control) on the protocol sheet.
  • Take microtiter strips out of the bag, take a minimum of one well as Blank. Store unused strips with desiccant at 2-8°C in the bag. Strips are stable until expiry date stated on the label.
  1. Add 50 µl STD/ SAMPLE/ CTRL (Standards/Sample/Control) in duplicate into respective well, except blank.
  2. Add 200 µl CONJ (Conjugate) into each well, except blank, swirl gently.
  3. Cover tightly and incubate for 3 hours at room temperature (12-26°C) in the dark.
  4. Aspirate and wash wells 5x with 300 µl diluted WASHBUF (Wash buffer), remove remaining WASHBUF by hitting plate against paper towel after the last wash.
  5. Add 200 µl SUB (Substrate) into each well.
  6. Incubate for 30 min at room temperature (18-26°C) in the dark.
  7. Add 50 µl STOP (Stop solution) into each well, shake well.
  8. Measure absorbance immediately at 450 nm with reference 630 nm, if available.

Typical Standard Curve

Manual

Product Manual


Publications

References

  • Utility of the Amino-Terminal Fragment of Pro Brain Natriuretic Peptide in Plasma. For the Evaluation of Cardiac Dysfunction in elderly Patients in Primary Health Care. Alehagen U et al., Clin Chem, 49:8; 1337-1346 (2003).
  • Head-to-head comparison of N-terminal pro-brain natriuretic peptide, brain natriuretic peptide and N-terminal pro-atrial natriuretic peptide in diagnosing left ventricular dysfunction. Hammerer-Lercher et al., Clin Chim Acta, 310(2),193-197 (2001).
  • Prognostic evaluation of neurohumoral plasma levels before and during beta-blocker therapy in advanced left ventricular dysfunction. Stanek B. et al., JACC, 38, 436-442 (2001).
  • Pulmonary arterial hypertension in rheumatic mitral stenosis: does it affect right ventricular function and outcome after mitral valve replacement? Pande S et al., Interact CardioVasc Thorac Surg, 9: 421-425 (2009).
  • Prognostic impact of body mass index in patients undergoing coronary artery bypass surgery. Sung SH et al., Heart, 97: 648-654 (2011).
  • Effect of piboserod, a 5-HT4 serotonin receptor antagonist, on left ventricular function in patients with symptomatic heart failure. Kjekshus JK et al., Eur J Heart Fail, 11: 771-778 (2009).
  • Plasma pro-B-type natriuretic peptide in the general population: screening for left ventricular hypertrophy and systolic dysfunction. Goetze JP et al., Eur Heart J, 27: 3004-3010 (2006).