The extraction and solubilization of proteins from seeds of Phaseolus vulgarisms for
two-dimension polyacrylamide gel electrophoresis (2D-PAGE) and analysis by mass
spectrometry are very sensitive procedures. In this study, we used two methods of
extraction and solubilization of proteins, the urea/thiourea method and the trichloroacetic
acid (TCA)/acetone precipitation method, in order to determine their effectiveness
in separating proteins from bean seeds by 2D-PAGE. In both methods, proteins
were well separated by 2D PAGE with minor variations in the protein pattern.
These two extraction methods showed that it was possible to separate hundreds of
very resolvent proteins by 2D electrophoresis. A protein spot was selected on the
2D-PAGE gel, digested with trypsin and analyzed by mass spectrometry (LCMS/
MS). The results suggest that thiourea/urea and TCA methods were effective and
reliable for the extraction and solubilization for 2D analysis of proteins from seeds of Phaseolus vulgaris .
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