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Two.dimensional separation of human plasma proteins using agarose gel isoelectric focusing followed by SDS capillary electrophoresis

Jun Mukai, Nao Yamaguchi and Takashi Manabe

CDepartment of Chemistry, Faculty of Science, Ehime University, Matsuyama, JAPAN

Abstract:

Two.dimensional separation of human plasma proteins combining agarose gel isoelectric focusing and SDS capillary electrophoresis was examined. After IEF, the IEF gel was dissected to 32 slices, then the proteins were extracted from the gel slices and were subjected to SDS.CE. Each electropherogram was converted into a density profile, the density profiles were arrayed in the order of IEF slice number and a 2.D density pattern was constructed. On the 2.D density pattern, about 90 protein “spots” were detected in pI range from 3.5 to 8.0 and molecular mass range from 20 kDa to 500 kDa. When the density pattern was compared with a pattern of 2.D gel electrophoresis, major protein spots could be correlated. This technique has the advantages in post.electrophoresis data processing when compared with conventional 2.D gel electrophoresis; 1) protein quantity can be obtained without staining, 2) molecular masses can be estimated from the migration time, 3) the separation results can be stored as digitized data.

Keywords: two−dimensional separation, agarose gel isoelectric focusing, plasma proteins, SDS capillary electrophoresis.

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