Enrichment of Lipoproteins by TX-114 Phase Partitioning
5:05
SDS-PAGE, Electroblotting, and Staining with Ponceau S
6:30
Tryptic Digestion, Lipopeptide Extraction from Nitrocellulose Membrane, Deposition onto MALDI Target, and Data Acquisition
8:47
Results: MALDI-TOF MS Analysis of N-terminal Lipopeptides
10:17
Conclusion
Transcript
The overall goal of this procedure is to extract lipoproteins from bacterial cells for direct applications and to further prepare N-terminal lipopeptides for structural analysis by MALDI-TOF mass spectrometry. This method can help answer key quest
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The enrichment of bacterial lipoproteins using a non-ionic surfactant phase partitioning method is described for direct use in TLR assays or other applications. Further steps are detailed to prepare N-terminal tryptic lipopeptides for structural characterization by mass spectrometry.