Isoelektrische Fokussierung in Mikrogelen zur Fraktionierung komplexer Proteingemische im Nanogrammbereich

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Abstract
A method is described for isoelectric focusing of complex protein mixtures in 2, 5 or 10 .mu.l capillaries. For 1 separation only 15-50 ng of a protein mixture is needed. Isoelectric focusing is finished after 10 min, and staining takes 20 min. Destaining takes approximately 30 min. Using defined mixtures of Servalyt from different pH ranges, isoelectric focusing can be adapted to the protein sample to be fractionated. Protein peaks separated by isoelectric focusing can be electrophoretically eluted and for further analysis refractionated directly in a microgradient gel. The resolution power of microisoelectric focusing is equal to the macroprocedure, as is demonstrated by isoelectric focusing of a protein test mixture, the water soluble proteins from cerebellum and heart, rat and human serum, and of a human oncocytoma of the thyroid gland.

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