QUANTITATIVE MEASUREMENT OF METAL ION CONCENTRATION OF PROTEINS SEPARATED BY ELECTROPHORESIS

This communication is devoted to nature determination and quantification by PIXE of metals contained in proteins after their separation by PolyAcrylamide Gel Electrophoresis (PAGE). After the electrophoresis, the gel is dried and each track is scanned with a 2.5 MeV proton beam which triggers metal X-ray fluorescence and then, allows to determine the type of metals contained in an electrophoretic band. For quantitative determination of the amount of the metal contained inside the band, the characteristic X-ray peak area is compared with those obtained with polyacrylamide gels doped with the same metal. The normalization has been achieved by using RBS measurements on the gel itself.

The procedure presented seems to be a very useful multielementary method for the metal content analysis and for the determination of the metal amounts inside proteins after their separation by electrophoresis. Furthermore it allows to check if metals remain bound to proteins.