2.9 Gel staining and analysis of protein synthesis

Two different stains, one with CoomassieBrillant Blue (CBB) for one-dimensional gels, and the second silver nitrate for two-dimensional gels, were used (this was prepared on the same gel after the first staining with CBB). In the case of CBB staining gels, they were incubated for 30 min in a dye solution containing 0.1% (w/v) Coomassie Blue - G250, 10% (v/v) acetic acid, and 25% (v/v) isopropanol and finally the gels were transferred to a bleaching solution of 10% (v/v) acetic acid and 10% (v/v) isopropanol. For silver nitrate staining, the suggested kit protocol was followed (ProteoSilver™ Plus, Sigma).

The image acquisition was performed by gel scanning III Image Scanner (GE Healthcare Life Sciences). Image Master 2D Platinum v 7.0 (GE Healthcare Life Sciences) software for comparative analysis of two-dimensional gels were used, which allowed to define differentially spots expressed between two or more situations analysed using the complimentary statistical analysis.