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Use of the PROTEAN Plus Dodeca Cell for Second-Dimension SDS-PAGE, Rev A

r constant voltage conditions, every gel experiences the same running conditions regardless of the number of gels in the tank. The temperature did not rise during the run and the buffer recirculation seemed to be particularly efficient in maintaining a uniform temperature throughout the inner buffer chamber.

Staining and Imaging
Post-electrophoresis, the gels were removed from the PROTEAN Plus hinged spacer plates. Care needs to be taken while removing 1.0 mm thick gels from their cassettes, as they are fragile and easily torn. Thicker gels would be easier to handle (Bio-Rads 1.5 mm cassettes are recommended, catalog #165-4171 for 20 x 20.5 cm and catalog #165-4174 for 25 x 20.5 cm cassette sizes). Ten of the gels were stained with colloidal Coomassie Blue for 48 hr and restained for a further 48 hr with fresh stain; one gel was stained with SYPRO Ruby protein gel stain (image not shown). The gels were destained with and stored in water. Gel images were acquired on an Agfa Arcus II scanner through a single thickness of red document wallet. The images were channel split in Microsoft Paintshop Pro software so that the red channel of the RGB TIFF file was selected.


Results and Discussion
Images of the 10 Coomassie Blue-stained gels run on the PROTEAN Plus Dodeca cell are shown in Figure 2. The gels are of excellent quality and are very consistent with the exception of some minor differences due to the isoelectric focusing step. The highly expressed proteins did not suffer from vertical streaking and the other proteins all show welldefined spots (see Figure 3). These high-quality gels are excellent for image analysis comparisons and were sufficiently loaded to a
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