Electrophoretic transfer of proteins from fixed and stained gels

A procedure by which sodium dodecyl sulfate gels can be fixed and stained with Coomassie blue and subsequently transferred to nitrocellulose for immunostaining is outlined. The procedure involves the complete removal of the stain followed by equilibration of the gel in sodium dodecyl sulfate running buffer. The efficiency of transfer is comparable to unfixed gels and the protein pattern of the transfer appears to be sharper, presumably due to less diffusion during the transfer process. The procedure does not affect the antigenicity of the proteins that have been examined by subsequent immunostaining. This method is particularly useful for situations in which sample size or concentration are limiting factors resulting in insufficient material for duplicate gels.