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.