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For Mr determinations, the protein samples were mixed 1:1 with 2¡Á SDS sample buffer containing ¦Â-mercaptoethanol and boiled for 5 min before being separated by
SDS-PAGE. Prestained Mr£¨·Ö×ÓÁ¿£© standards (BioRad) were run on each gel as a standard. Following electrophoresis, proteins were either visualized with Commassie Blue dye (BioRad) or transblotted to PVDF membrane (BioRad) using a transblot semidry electrophoretic transfer cell at 20V for 25¨C30 min. Transfers were done
using 48 mM Tris, 39 mM Gly, 0.0375% (w/v) SDS, and 12% (v/v) methanol (pH 9.2) as a transfer buffer. Immunoblots were incubated with a 1:100 dilution (diluted in 1% BSA/TBST) of PPO antiserum developed from our purified wheat bran protein.Antigens were visualized by alkaline phosphatase-conjugated goat and antimouse (Sigma) and 5-bromo-4-chloro-3-indolyl phosphate and nitro blue tetrazolium (Sigma).

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