Archived polyacrylamide gels as a resource for proteome characterization by mass spectrometry |
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Authors: | Shevchenko A Loboda A Ens W Schraven B Standing K G Shevchenko A |
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Institution: | Peptide and Protein Group, European Molecular Biology Laboratory, Heidelberg, Germany. schevchenko@EMBL-Heidelberg.de |
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Abstract: | Mass spectrometry was applied to identify protein spots excised from an archived two-dimensional polyacrylamide gel that had been dried and stored for eight years at room temperature. All proteins were successfully identified. Detailed characterization of protein digests by matrix-assisted laser desorption/ionization (MALDI) peptide mapping, nanoelectrospray tandem mass spectrometry and MALDI-quadrupole time-of-flight mass spectrometry revealed no evidence of protein degradation or modifications that could hamper identification of proteins in a sequence database. The experiment with a model protein demonstrated that the pattern of tryptic peptides and the yield of individual peptides were not noticeably changed in the in-gel digest of the archived protein spot compared to the digest of the spot excised from a fresh gel. Thus, the characterization of "archived proteomes" has the potential to advance proteomic research without repeating "wet" biochemistry experiments, that had been perfected in the laboratory years ago. |
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