Planar Capillary Electrophoresis on Slab Gels

Prior to the development of CZE, the primary application of electrophoresis was for large biological macromolecule separations on planar gels. The gels were supported on glass plates not unlike the particles in the thin layer of a TLC plate. These so-called slab gels were needed to confine the separation buffer. It was easier to prepare cross-linked 

After spotting After 1st develop. After 2nd develop. 

The 2D-SDS-PAGE separation described results in a planar array of separated compounds. If these are unknown proteins (perhaps just one or a few which interrupt a standard pattern, and may signal a mutant form or an abnormal biological state), the spots of interest may be cut out or dissolved out of the gel. The isolated spot can be subjected to tryptic digest, the peptide digest mixture separated by capillary HPLC, and the separated peaks characterized, and if necessary, sequenced by ESI-MS and ESI-MS-MS, as was illustrated in Figs. 13.21 and 13.22. If the 2D separation is of peptides from a single protein in a 

Berger, C., Ed. Supercritical Fluid Chromatography Marcel Dekker, Inc. New York, 1998. Ardrey, R.E. Liquid Chromatography—Mass Spectrometry, An Introduction Wiley New York, 2003. Berthold, et al. Micellar Liquid Chromatography Marcel Dekker, Inc. New York, 2000. Camilleri, P. Capillary Electrophoresis Theory and Practice CRC Press Boca Raton, EL, 1998. Cazes, J. Encyclopedia of Chromatography Marcel Dekker Inc. New York, 2001. 

Cunico, R.L. et al. Basic HPLC and CE of Biomolecules Bay Anal3dical Laboratories, 1998. Fritz, J.S. Gjerde, D.T. Ion Chromatography VCH Publishers Germany, 2000. 

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