Coomassie colloidal blue g 250

Gels were fixed for about 8 h in solution (10% (v/v) acetic acid, 40% (v/v) ethanol, and 50% (v/v) water), washed 3 times in water, and then stained with Coomassie colloidal blue G-250 according to the GE handbook (GE Healthcare) with minor modifications. Gel images were acquired with the PowerLook 2100XL color scanner (UMAX Technologies, Atlanta, CA, USA) at a resolution of 16 bits and 300 dpi and were assayed by Image master 2D Platinum Software Version 6.0 (GE Healthcare). To compare the spot quantities between gels accurately, each spot volume was normalized as a percentage of the total volume of all of the spots in the gel. All automatic spot detections in each gel were manually inspected and edited as necessary to confirm the absence of mismatched and unmatched spots. Differentially expressed protein spots were changed abundance by at least ±1.2-fold, with an error probability of p < 0.05.

Gels were fixed for about 8 h in a solution containing (10% (v/v) acetic acid, 40% (v/v) ethanol, and 50% (v/v) water), washed three times in water, and then stained with Coomassie colloidal blue G-250 according to the GE handbook (GE Healthcare) with minor modifications. Gel images were acquired with the PowerLook 2100XL color scanner (UMAX Technologies, Atlanta, CA, USA) at a resolution of 16 bits and 300 dpi, and were assayed by Image master 2D Platinum Software Version 6.0 (GE Healthcare). To limit experimental variation among 2D gels, quantitative comparison of protein spots was performed on the base of their percentage volumes. All automatic spot detections for each gel were manually inspected and edited as necessary to confirm the absence of mismatched and unmatched spots. One-way ANOVA and comparison of treatment means were carried out on the SAS program. Differentially expressed protein spots were (1) consistently present in all replicates and (2) changed abundance by at least ±1.2-fold, with an error probability of P≤0.05.