Development of a New Type of Two-Dimensional Electrophoresis and Its Application to the Analysis of Alkaline Phosphatase Isoenzymes in Sera of Pregnant Woman

新しい2次元電気泳動法の開発と本法を用いた妊婦血清中アルカリホスファターゼアイソザイムの分析

Development of a New Type of Two-Dimensional Electrophoresis and Its Application to the Analysis of Alkaline Phosphatase Isoenzymes in Sera of Pregnant Woman

Creators Hirota Masayuki

1) The author developed a highly sensitive staining method of alkaline phosphatase (ALP) by using an indigo derivative (TBIP) as the substrate together with an accelerator tetrazolium (NBT). By this system, 2x10^<-5> IU of ALP activity or ALP activity in 1 μl normal serum can be demonstrated as a distinctive violet band. This staining method can also be coupled with the CBBG-protein staining. By this double staining, each ALP isoenzymes can be easily identified in relation to the localization of some known serum proteins on the protein-map. 2) The author developed a new type of two-dimensional PAG electrophoresis (2-DE). The first electrophoresis is capillary isoelectric focusing of 6% PAG, in pH range 3.5-10. The second electrophoresis is 12.5% PAG slab electrophoresis using Cl^-/glycinate^- stacking system. The procedure of this 2-DE system is simple and sensitive enough to sharply separate ALP isoenzymes in the liver or placenta.

[ISSN]0513-1731

[NCID]AN00243156