MICROPREPARATIVE ONE-DIMENSIONAL AND 2-DIMENSIONAL ELECTROPHORESIS - IMPROVEMENT WITH NEW PHOTOPOLYMERIZATION SYSTEMS

To improve the efficiency of one- and two-dimensional electrophoresis for micropreparative purposes, the use of gels polymerized with other initiators than the standard N,N,N',N'-tetramethylethylenediamine (TEM ED)/persulfate systems for sodium dodecyl sulfate electrophoresis has been investigated. We show here that the recently described photoiniti ator system, composed of methylene blue, toluene sulfinate and dipheny liodonium chloride, leads to a decreased resolution. Resolution can be restored if methylene blue is replaced by riboflavin. Two-dimensional electrophoresis with mg loadings of proteins has also been evaluated with these systems. Independently of the polymerization system, resolu tion for the first dimension is low with rod gels, increases with gel strips and is further improved when immobilized pH gradients are used. Here too, only the riboflavin/sulfinate/iodonium system results in a resolution that matches the one obtained with the standard TEMED/persu lfate system. Gels polymerized with the riboflavin/sulfinate/iodonium system yield better results upon N-terminal microsequencing after blot ting than gels polymerized with the standard TEMED/persulfate system.