Mouse IL-17A Culture media OneStep ELISA Kit

ELISA

• Standard Curve
  Example of Mouse NT-pro BNP standard curve in Assay Diluent GA2. Plotted are the background-subtracted data.
  This standard curve is provided for demonstration only.

• Spike Recovery
  The recovery of Mouse IL-17A was evaluated in activated samples spiked with concentrations spanning the entire assay range.

• Dilution Linearity
  The concentrations of Mouse IL-17A were measured and interpolated from the target standard curves and corrected for sample dilution.
  The leftmost bar in the graph shows biological samples diluted in Assay Diluent to 0.1% concentration. "undiluted" was defined as 0.1% biological sample. The subsequent bars represent two-fold serial dilutions of this reference. The mean target concentration was determined to be 769.48 ng/mL in mouse EL-4 cells culture supernatant stimulated with 10 ug/mL PHA and 10 ug/mL PMA for 2 days.

• Dilution Linearity
  The concentrations of Mouse IL-17A were measured and interpolated from the target standard curves and corrected for sample dilution.
  The leftmost bar in the graph shows biological samples diluted in Assay Diluent to 10% concentration. "undiluted" was defined as 10% biological sample. The subsequent bars represent two-fold serial dilutions of this reference. The mean target concentration was determined to be 3118.84 pg/mL in unstimulated mouse EL-4 cells culture supernatant.

• Dilution Linearity
  The concentrations of Mouse IL-17A were measured and interpolated from the target standard curves and corrected for sample dilution.
  The leftmost bar in the graph shows biological samples diluted in Assay Diluent to 25% concentration. "undiluted" was defined as 25% biological sample. The subsequent bars represent two-fold serial dilutions of this reference. The mean target concentration was determined to be 1440.13 pg/mL in mouse lung tissue culture supernatant stimulated with 5 μg/mL lipopolysaccharide for 6 days.

• Spike-and-dilution Linearity
  To evaluate assay linearity, the sample was spiked with high levels of Mouse IL-17A 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, 24 replicate measurements of the diluent (zero calibrator) were performed. The mean (AVERAGE) and standard deviation (STDEV) of the 24 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 IL-17A and Human IL-17A 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 IL-17A was tested in the Mouse IL-17A 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