NT-proBNP ELISA Assay Kit


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

NT-proBNP ELISA Assay Kit

NT-proBNP ELISA Assay Kit Developed and Manufactured in Austria by Biomedica

Size: 1×96 wells
Sensitivity: 3 pmol/L
Dynamic Range: 0 – 640 pmol/L
Incubation Time: 3.5 hours
Sample Type: Serum, Saliva
Sample Size: 50 µL
Species: Human
Alternative Names: Brain Natriuretic Peptide ELISA, N-Terminal ProBNP ELISA
For Research Use Only

Controls Included

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.

Related Products

BNP Fragment ELISA Kit
NT-proANP ELISA Assay Kit
NT-proCNP ELISA Assay Kit

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

Additional Information

Assay Principle

This kit is a sandwich enzyme immunoassay for the quantitative determination of NT-proBNP in human serum and EDTA plasma. In a first step, sample and conjugate (sheep anti-human NT-proBNP-HRP) are pipetted into the wells of the microtiter strips, which are pre-coated with polyclonal sheep anti-NT-proBNP antibody. NT-proBNP present in the sample binds to the pre-coated antibody in the well and forms a sandwich with the conjugate (detection antibody). In the washing step all non-specific unbound material is removed. In a second step, the substrate (Tetramethylbenzidine; TMB) is pipetted into the wells. The enzyme catalyzed color change of the substrate is directly proportional to the amount of NT-proBNP present in the sample. This color change is detectable with a standard microtiter plate ELISA reader.

The NT-proBNP ELISA kit uses highly purified, epitope-mapped antibodies. The antibodies utilized in the NT-proBNP ELISA (BI-20892) are as follows:

Capture antibody: AA 32-57 (polyclonal sheep anti-human NT-proBNP)
Detection antibody: AA 8-29 (HRP-labeled polyclonal sheep anti-human NT-proBNP)

Assay Procedure

  • 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

NT-proBNP ELISA Standard Curve

Package Inserts

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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