Abstract
MALDI-TOF mass spectrometers are now commonplace and their relative ease of use means that most non-specialist labs can readily access the technology for the rapid and sensitive analysis of biomolecules. One of the main uses of MALDI-TOF-MS is in the identification of proteins, by peptide mass fingerprinting (PMF). Here we describe a simple protocol that can be performed in a standard biochemistry laboratory, whereby proteins separated by 1D or 2D gel electrophoresis can be identified at femtomole levels. The procedure involves excision of the spot or band from the gel, washing and destaining, reduction and alkylation, in-gel trypsin digestion, MALDI-TOF-MS of the tryptic peptides and database searching of the PMF data. Up to 96 protein samples can easily be manually processed at one time by this method.
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Acknowledgments
The authors acknowledge the support of the Biotechnology and Biological Sciences Research Council, UK.
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© 2012 Springer Science+Business Media, LLC
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Webster, J., Oxley, D. (2012). Protein Identification by MALDI-TOF Mass Spectrometry. In: Zanders, E. (eds) Chemical Genomics and Proteomics. Methods in Molecular Biology, vol 800. Humana Press. https://doi.org/10.1007/978-1-61779-349-3_15
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DOI: https://doi.org/10.1007/978-1-61779-349-3_15
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Publisher Name: Humana Press
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Online ISBN: 978-1-61779-349-3
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