Standard Curve
Example of Mouse NT-Pro BNP standard curve in Assay Diluent HS4. Plotted are the background-subtracted data.
This standard curve is provided for demonstration only.
Mouse NT-Pro BNP OneStep ELISA Kit
Mouse NT-Pro BNP OneStep ELISA Kit
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Product Details
Product Details
Product Specification
| Antigen | NT-pro BNP |
| Immunogen | Recombinant Protein |
| Antibody Type | Recombinant mAb |
| Reactivity | Ms |
| Purification | Protein A |
| Stability & Storage | 12 months from date of receipt / reconstitution, 2 to 8°C as supplied. |
Kit
| Precision | Intra-assay: 3.0%； Inter-assay: 4.7% |
| Sample type | Serum; EDTA Plasma, Heparin Plasma, Citrate Plasma; Cell Culture Supernatant |
| Assay type | Sandwich (quantitative) |
| Sensitivity | 2.49 pg/mL |
| Range | 4.69 pg/mL – 300 pg/mL |
| Recovery | Serum: 98% EDTA Plasma: 104% Citrate plasma: 91% Heparin Plasma: 102% Cell Culture Supernatant: 115% |
| Assay time | 60 minutes |
| Species reactivity | Ms |
Background
Mouse NT-Pro BNP (N-terminal pro-brain natriuretic peptide) is a 76-amino acid polypeptide produced by cardiac myocytes, primarily in the ventricles. It is an inactive cleavage fragment derived from the precursor Pro BNP during the processing to mature BNP. Unlike the active BNP hormone, NT-Pro BNP has no known biological function and is cleared more slowly from the circulation. Its secretion increases significantly in response to cardiac wall stretch, pressure overload, and heart failure. In mouse models, elevated NT-Pro BNP levels serve as a reliable biomarker for myocardial stress and ventricular dysfunction. Researchers commonly measure Mouse NT-Pro BNP to assess the severity of cardiac conditions such as hypertrophy and cardiomyopathy. Because it remains stable in blood samples, NT-Pro BNP is a practical and sensitive indicator for preclinical heart disease studies. Thus, it plays a valuable role in evaluating therapeutic interventions in murine models of heart failure.
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ELISA
Spike Recovery
The recovery of Mouse NT-Pro BNP was evaluated in activated samples spiked with concentrations spanning the entire assay range.
Dilution linearity
The concentrations of Mouse NT-Pro BNP were measured and interpolated from the target standard curves and corrected for sample dilution.
The leftmost bar in the graph represents neat biological samples. "Undiluted" was defined as the neat biological sample (100%). The subsequent bars represent two-fold serial dilutions of this reference. The mean target concentration was determined to be 36.6 pg/mL in Mouse heart tissue culture supernatant.
Spike-and-dilution Linearity
To evaluate assay linearity, three samples were spiked with high levels of Mouse NT-Pro BNP in different matrices and serially diluted with the corresponding Calibrator Diluent to fall within the assay's dynamic range.
Intra-Assay Precision (Precision within an assay)
Three samples of known concentration were tested fifteen times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays).
Three samples of known concentration were tested in separate assays to assess inter-assay precision. Assays were performed with at least three lots of components.
Determination of Minimum Detectable Dose (MDD)
The MDD was determined using three independent lots of assay components. For each lot, 19 replicate measurements of the diluent (zero calibrator) were performed. The mean (AVERAGE) and standard deviation (STDEV) of the 19 replicates were calculated. The MDD for each lot was then calculated according to the following formula:
MDD = 2 × STDEV + AVERAGE
Cross-reactivity
The cross-reactivity between Mouse BNP and Human BNP was assessed by testing both proteins in the same assay system. Serial dilutions of each protein were measured, and the dose-response curves were compared. No significant cross-reactivity was observed when Human NT-pro BNP was tested in the Mouse NT-Pro BNP assay, as the signals remained within the background range of the assay.
HOOK Effect Threshold
The upper limit of the HOOK effect was established at 100× the highest calibrator concentration (equivalent to a 2-log10 increase). No HOOK effect was observed below this threshold, confirming that samples within this concentration range yield reliable quantitative results without signal depression.
Protocol Diagram
