Mass spectrometric characterization of proteins transferred from polyacrylamide gels to membrane filters |
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Authors: | Ino Yoko Hirano Hisashi |
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Institution: | Advanced Medical Research Center/Graduate School of Nanobioscience, Yokohama City University, Kanazawa, Yokohama, Japan. |
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Abstract: | In the 1990s, a technique was developed to transfer proteins from electrophoresis gels onto poly(vinylidene difluoride) (PVDF) membranes, digest the proteins on the membranes with proteases such as trypsin and analyze the resulting peptides on the membranes directly by mass spectrometry to identify the proteins. This technique, based on gel electrophoresis, is particularly useful for analyzing protein isoforms, splicing variants and post-translationally modified proteins. Previously, the low ionization efficiency of peptides immobilized on the membranes often rendered this technique useless. However, this technique has been improved by the use of PVDF membranes with a small pore size, which has enabled highly efficient and effective electroblotting and mass spectrometric analyses. Here, the advantage of this technique is discussed. |
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