ELISA inhibition
Details of the ELISA method were described in a previous report.11 Pierce™ Protein-Free blocking buffer (Thermo Fisher Scientific, Uppsala, Sweden) was used for blocking and serum dilution. Proteins were extracted from wheat and barley using a Mammalian cell lysis kit (Sigma-Aldrich, Missouri, America) and added to 2% serum or blocking buffer at a concentration of 0, 0.01, 0.1, 1, and 2 mg/mL and incubated for 1 h at 25°C, and then used for the assay. The inhibition rate was calculated as a ratio to the measured value when no protein was added to the serum.