All samples were subjected to SDS-polyacrylamide gel
electrophoresis (SDSseparated according to the method of Laemmli (1970). The
separation was performed with a 10% separating gel and a 5%
stacking gel using the miniVE Vertical electrophoresis system (GE
Healthcare, U.S.A). The relative molecular mass (Mr) of each
protein was determined using a standard curve generated from
standard set of 12 pre-stained proteins in the range 3.5-260 kDa
(Novex Sharp Protein Standard, Invitrogen, USA).
The gels were visualized by staining with Colloidal Coomassie
Brilliant Blue G-250 (CBB) as described by Neuhoff et al. 1988.
After gel electrophoresis, the gel was transferred into a fixative
solution (50% ethanol, 2% Phosphoric acid in ddH2O) for 1 h at
room temp. The gel was stained with CBB (0.2% w/v) for 12-24 h by
agitation on a shaker overnight. The staining solution was then
removed. The gel was destained with several changes of ddH2O.