2.5. Procedure of FLISA

The procedures for FLISA and ic-ELISA were identical before the secondary antibody was added. More specifically, after washing with PBST for three times, 100 μL of 655 nm QDs-conjugated goat anti-mouse IgG (1:10,000 dilution) was added to each well and incubated at 37 °C for 45 min. After three washes with PBST, a SpectraMax M2e Microplate Reader (Molecular Devices, San Jose, CA, USA) was used to measure the fluorescence signals from samples with excitation/emission at 360/655 nm. The fluorescence intensities were identified as B₀ and B for the control and testing sample. The standard curve was established by plotting the form (B/B₀) 100% against the 2-NP-AMOZ concentration.