Polyacrylamide gel electrophoresis (PAGE)
Molecular weights of polypeptides in defatted silflower, freeze-dried
soluble protein fractions, and spent material after sequential
extraction of soluble proteins were estimated by gel electrophoresis.
Native gel electrophoresis was done according to the method reported
previously by Hojilla-Evangelista and Evangelista (2006), with these
modifications: 1) sample concentrations were 10 mg/mL (quantity basis),
2) samples dissipated undisturbed in the native sample buffer for 1 h at
room temperature before being centrifuged at 11,000 rpm for 3 min, and
3) loading volume was 15 µL.
SDS-PAGE was done on the reduced proteins from the same samples used for
native gel electrophoresis following the method of Hojilla-Evangelista
et al. (2013), with slight modifications: sample concentrations were 5
mg/mL, protein standard was BioRad Precision Plus Dual Xtra (molecular
weight range 2 to 250 kDa) and load volume for samples was 15 µL.