In-gel digestion of proteins for MALDI-MS fingerprint mapping

Current Protocols in Protein Science
C R JiménezA L Burlingame

Abstract

Mass spectrometry (MS) has emerged as a sensitive, versatile, and rapid method for protein identification, following the advent of electrospray ionization mass spectrometry (ESI-MS) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The advantages of MALDI-MS over ESI-MS include its relatively high tolerance to contamination from biological matrices, its high sensitivity, the relative ease of interpreting spectra from mixtures, and the formation of singly protonated molecular ions for tandem analysis. Peptide fingerprint mass mapping and partial peptide sequencing using post-source decay and ladder sequencing by MALDI-MS in combination with algorithms for sequence database interrogation have the potential for identification and structural investigation of proteins. This unit describes in-gel digestion for peptide mass mapping of picomole to subpicomole quantities of protein derived from Coomassie- or silver-stained polyacrylamide gels. After digestion, the peptides are extracted from the gel and mass analyzed.

References

Oct 1, 1995·Electrophoresis·S D Patterson, R Aebersold
Mar 1, 1996·Analytical Chemistry·A ShevchenkoM Mann
Nov 14, 1997·Analytical Chemistry·C Fenselau
May 1, 1997·Mass Spectrometry Reviews·P Jungblut, B Thiede

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Citations

Jul 17, 2012·Applied and Environmental Microbiology·Zachary L PrattCharles W Kaspar
Mar 29, 2014·Biochemical and Biophysical Research Communications· VikrantAshok K Varma
Feb 26, 2010·Nature·David K BreslowJonathan S Weissman
Aug 12, 2014·Journal of Biomolecular Structure & Dynamics· VikrantAshok K Varma
Oct 22, 2009·Letters in Applied Microbiology·C D MillerA J Anderson
Jun 27, 2012·Protein Science : a Publication of the Protein Society·Youngil ChangChiwook Park
Dec 29, 2010·Plant Molecular Biology·Hong YaoJames A Birchler
May 1, 2011·Current Protocols in Chemical Biology·Jacob S YountHoward C Hang
Dec 12, 2001·Biochimica Et Biophysica Acta·M D StubbsG B Moorhead

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