Miniprotein 3 unit

Aliquots taken at different digestion time points were evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which was conducted at a presence of 5% β-mercaptoethanol by using a 12% polyacrylamide gel. The gels were stained with 0.1% (w/v) Coomassie brilliant blue R-250 (Sigma) at room temperature. SDS-PAGE was conducted on a Bio-Rad Miniprotein 3 unit (Bio-Rad Laboratories, Inc., Hercules, CA, USA) with voltage 60 V for stacking gel, and followed by 120 V for separating gel. The molecular weight values of the protein fractions were estimated using broad-range Protein Marker (15–150 kDa). The protein-stained bands were scanned with Image Scanner III (GE Healthcare Biosciences, Uppsala, Sweden) and analyzed using Quantity One software, version 4.6.2 (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was employed to analyze protein profiles of glycinin during fermentation. SDS-PAGE was carried out in a Bio-Rad Miniprotein 3 unit (Bio-Rad Laboratories, Inc., Hercules, CA, USA) with stacking gel at 4% and separating gel at 12%. SDS-PAGE was carried out under reducing conditions by supplying 5% β-ME. Running the gel at 80 V and 120 V for stacking and separating gel, respectively. A pre-stained broad range standard (15–130 kDa) was used. Gels were then scanned with Image Scanner III (GE Healthcare Biosciences, Uppsala, Sweden) and analyzed by using Quantity One software, version 4.6.2 (Bio-Rad Laboratories, Inc., Hercules, CA, USA).