Unit

UNIT 16.4 In-Gel Digestion of Proteins for MALDI-MS Fingerprint Mapping

  1. C.R. Jiménez,
  2. L. Huang,
  3. Y. Qiu,
  4. A.L. Burlingame

Published Online: 1 MAY 2001

DOI: 10.1002/0471140864.ps1604s14

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Jiménez, C., Huang, L., Qiu, Y. and Burlingame, A. 2001. In-Gel Digestion of Proteins for MALDI-MS Fingerprint Mapping. Current Protocols in Protein Science. 14:16.4:16.4.1–16.4.5.

Author Information

  1. University of California, San Francisco, San Francisco, California

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: DEC 1998

This is not the most recent version of the article. View current version (1 NOV 2016)

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.