2-DIMENSIONAL ELECTROPHORESIS - AGAROSE-GEL ISOTACHOPHORESIS FOLLOWED BY SODIUM DODECYL-SULFATE POLYACRYLAMIDE ELECTROPHORESIS

High resolution of proteins by gel isotachophoresis (ITP) has been achieved in previous work, and two-dimensional electrophoresis was applied in order to obtain further information on the proteins separated by ITP in agarose gels. Gradient or discontinuous buffer acrylamide gels in the presence of sodium dodecyl sulphate (SDS) and beta-mercapthoethanol were used for the second electrophoresis. After two-dimensional ITP-SDS polyacrylamide gel electrophoresis (PAGE), several components were observed on the plates after either silver or Coomassie Brilliant Blue staining. The enhanced technique with SDS-PAGE revealed further heterogeneity among C3-complement proteins, with at least eight alpha- and eight beta-chains of C3-complement. Two albumins spots were also observed after staining. The proposed technique provides a powerful addition to earlier methods for the separation of proteins.