Figure 5

Effects of FIGE on protein separation under native gel conditions. Comparison of GroEL 14-mer complex (840 kDa) (a) or MCF-7 cell nuclear extract (b) in native PAG between CFE and FIGE conditions. FIGE was the left panel. CFE control was the right panel. The gels were 1 mm × 7 cm native 6% PAG casted in a mini-Protean 3 apparatus. Run time was 2 hrs 15 min in control condition. Pulsed condition run time was 5 hrs 30 min with a t a/t r of 400/100 msec. Lane 1, native MW markers (10 μg total); Lane 2, 10 μg purified E. coli GroEL native complex (14-mer, 840 kDa). The band at 300 kDa could be a minor cofactor associated with GroEL during purification. The gel was stained with Coomassie blue. Proteins were purposefully overloaded to best represent the effect of "detrapping" of native proteins under pulsing conditions. In panel b), MCF-7 nuclear extracted were run through native 2–5% PAG. The run time was 10 hrs for CFE control condition. Pulsed condition run time was 14 hrs with ta/tr of 900 msec/240 msec. Lane 1, 5.6 μg MCF-7 cell nuclear extract; Lane 2, 7.5 μg MCF-7 cell nuclear extract; Lane 3, native protein MW markers. The gels were silver stained.