Immunoblotting and immunoblotting inhibition
Two-dimensional (2D) polyacrylamide gel electrophoresis (PAGE) and immunoblotting were performed by slightly modifying previously reported methods.12 A total of 25 μg of the extracted protein was subjected to immunoblotting, and Pierce ™ Protein-Free blocking buffer was used for blocking and serum dilution. The proteins extracted from wheat and barley were added to 2% serum or blocking buffer at a concentration of 0, 0.1, and 2 mg/mL, incubated for 1 h at 25°C, and then used for immunoblotting.