Human NT-proBNP ELISA Kit

Catalog No.: BEK1178

Size: 96T

Range: 62.5 pg/ml - 4000 pg/ml

Sensitivity: 2 pg/ml

Storage and Expiration: Store at 4℃ for 6 months, or at -20℃ for one year. Store HRP conjugated antibody at 4℃ for one year.

Application: For quantitative detection of human NT-proBNP in serum, plasma and cell culture supernatants.

Introduction

The N-terminal prohormone of brain natriuretic peptide (NT-proBNP) is a 76 amino acid N-terminal fragment of brain natriuretic peptide. Both BNP and NT-proBNP levels in the blood are used for screening, diagnosis of acute congestive heart failure (CHF) and may be useful to establish prognosis in heart failure, as both markers are typically higher in patients with worse outcome.BNP may be a useful screening tool for left ventricular dysfunction in patients with history suggestive of heart disease and be used to assist in forming a pretest probability, which in turn could greatly assist in appropriateness of patient referral and in optimization of drug therapy. 

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-NT-proBNP polyclonal antibody was pre-coated onto 96-well plates. The standards and test samples were added to the wells and any NT-proBNP present is bound by the immobilized antibody. After washing away any unbound substances, the HRP conjugated anti-NT-proBNP monoclonal antibody was added to wells as detection antibodies. Following a wash to remove any unbound antibody-enzyme reagent, the TMB substrates were added to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding stop solution. The density of yellow is proportional to the NT-proBNP amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of NT-proBNP can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human NT-proBNP antibody
2. Human NT-proBNP standards: 2 tubes (10 ng/ tube)                  
3. Sample / Standard diluent buffer: 30ml
4. HRP conjugated anti-human NT-proBNP antibody (Concentrated): 130 μl. Dilution: 1:100 (Store at 4℃, do NOT store it at -20℃)
5. Antibody diluent buffer: 12 ml
6. TMB substrate: 10 ml
7. Stop solution: 10 ml
8. Wash buffer: 30 ml (25x). Dilution: 1:25

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. Microplate reader (wavelength: 450nm)
2. Precise pipette and disposable pipette tips
3. Deionized or distilled water
4. Automated plate washer
5. Beakers, flasks or cylinders, etc.
6. Statistical calculator
7. Plate cover
8. Absorbent filter papers or absorbent material
9. ELISA shaker