Rice Protein Extraction and SDS-PAGE

A previous report outlined the process of protein extraction from rice powder and the method of moderate denaturation to achieve higher resolution SDS-PAGE results (Takahashi et al. 2019 (link)). SDS-PAGE was performed using 15% polyacrylamide gels (ATTO, Tokyo, Japan) with protein-size markers (Bio-Rad, Hercules, MA, USA). After separation, the proteins were stained with 0.1% Coomassie Brilliant Blue (CBB) R-250 (Nacalai tesque, Kyoto, Japan). The gel was destained for at least 3 h before being scanned directly. Quantity One software ver. 4.6.9. (Bio-Rad) was used to determine the intensity of TP intensity in a lane, glutelin acidic subunit, glutelin basic subunit, and prolamin. The ratios of glutelin (G) / TP, prolamin (P) / TP, and other proteins / TP were calculated as follows: