Figure 1.

Optimization of a luminol-based chemiluminescence assay using a luminometer. Various concentrations of H₂O₂ (0, 1, 5, and 10 μM), luminol (25, 50, 100, and 200 μM in NaOH), and 4IPBA (0 or 180 μM) were added to a black-walled glass bottom 96-well plate (see Table 1). Chemiluminescence was measured using a luminometer with either (a) 0.1 unit/ml or (b) 0.2 unit/ml HRP for the indicated times (n = 3, ±SD). (c) Shown are R ² values for different buffer conditions. R ²: linear correlation coefficient.