Spots can be excised from fluorescently stained gels using a KBiosystems KCore. The resulting spots are digested robotically with trypsin using a Genomic Solutions ProGest. The digests are cleaned by Ziptip using a Packard Multiprobe IIEX. Peptides and the matrix are spotted automatically to 4700 target plates using the Applied Biosystems Symbiot XVI. A schematic depiction of this process provides a complete overview of this method.
Example of Process Workflow for Pyruvate Kinase:
A Peptide Mass Fingerprint from a Pseudomonas syringae protein.
| The five peaks with red arrows have a MSMS image displayed below in order from largest m/z to smallest. |
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| MSMS of m/z 2925 from PMF: |
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| MSMS of m/z 2762 from PMF: |
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| MSMS of m/z 1827 from PMF: |
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| MSMS of m/z 1599 from PMF: |
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| MSMS of m/z 1258 from PMF: |
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Once the data is submitted for a search, a MASCOT search result is obtained.