Rehydration of polyacrylamide gels

A variety of methods have been published over the past decade, but the most popular versions involve washing any residual SDS from the stained gel pieces with 50% acetonitrile followed by desiccation. The dried gel pieces are rehydrated with an initial aliquot of enzyme followed by further additions of buffer as the gel pieces re-swell, drawing the protease further into the polyacrylamide. Then, after a suitable incubation interval, the resultant peptides are leached from the gel pieces with 60% acetonitrile, concentrated, and applied to HPLC separation. 

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