A novel PVDF membrane for on-membrane identification of gel-resolved proteins by mass spectrometry

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In the 1990s, techniques were developed to transfer proteins from gels onto PVDF membranes, and to identify the proteins on the membranes by MALDI-MS. However, the low ionization efficiency of peptides and proteins immobilized on the membranes often renders these techniques useless. Therefore, it is essential to develop membranes on which peptides and proteins can be efficiently ionized for MS analysis. In this study, we found that 0.1-μm pores PVDF membranes can be used for highly efficient and effective electroblotting and MALDI-MS analysis. This membrane showed high ion yield in MALDI-MS analysis of peptides compared to conventional PVDF membranes. In all, 38 yeast proteins with various physicochemical characteristics were separated by 2-DE, immobilized onto the 0.1-μm pore PVDF membranes, and successfully identified by MALDI-MS/MS.