Preparative Electroelution of Proteins from Polyacrylamide Gels

1 Preparative Electroelution of Proteins from Polyacrylamide Gels 

This protocol is designed for proteins separated by SDS-containing PAGE systems. When proteins shall be eluted from gels of other systems, the respective electrode buffer has to be used instead of Soln. A and the polarity should be taken into account. 

The content of glycerol or sucrose in Soln. A complicates the removal of SDS therefore, the sample has to be dialyzed after electroelution. To avoid difficulties use Soln. A without glycerol or sucrose. 

20 mM ammonium hydrogencarbonate buffer, pH 7.5 contains maximal 0.05% SDS if necessary 80% TCA (w/v) in ddH20 1 mM HCl in acetone Acetone 

The gel slices containing the protein of interest are minced in the presence of Soln. A (buffer volume nearly the same of gel volume), filled into a vessel of appropriate size and sonicated for 30 - 60 min. 

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