Fig 4.

Continuous elution SDS–10% PAGE using a Model 491 Prep Cell. Whole-cell lysates of WT CO92 were run on the gel, and the fractions were collected. Every four fractions were pooled, concentrated, dialyzed, subjected to SDS–12% PAGE, and stained with SYPRO-Ruby (left panel). An aliquot of the starting crude material was also subjected to electrophoresis. A parallel gel was subjected to Western blot analysis using hyperimmune rat sera (Fig. 1B) (right panel). The Western blot bands that corresponded to the SYPRO-Ruby-stained protein bands were picked and subjected to MALDI-TOF for identification. The identity of spots is indicated. The presence of multiple bands on the Western blot represented either aggregated or degraded version of the same proteins, as confirmed by mass spectrometry of several bands. The migration of molecular mass markers is also shown.