Silver stain removal using H 2 O 2 for enhanced peptide mass mapping by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry

Hydrogen peroxide solutions are reported for the removal of silver stain from proteins isolated in polyacrylamide gels. Removal of silver stain prior to in‐gel digestion is shown to enhance sensitivity and sequence coverage of matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrome...

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Bibliographic Details
Published in:Rapid communications in mass spectrometry 2002-02, Vol.16 (3), p.160-168
Main Authors: Sumner, Lloyd W., Wolf‐Sumner, Barbara, White, Steven P., Asirvatham, Victor S.
Format: Article
Language:English
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Summary:Hydrogen peroxide solutions are reported for the removal of silver stain from proteins isolated in polyacrylamide gels. Removal of silver stain prior to in‐gel digestion is shown to enhance sensitivity and sequence coverage of matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOFMS) peptide mass maps. The rate of silver removal using H 2 O 2 is influenced by H 2 O 2 concentration and increases with increasing pH. The presence of complexation reagents such as ammonia from mass spectrometry compatible ammonium bicarbonate solutions enhances the efficiency and speed of H 2 O 2 ‐mediated silver removal. H 2 O 2 ‐mediated silver removal using the described procedure does not appear to have any detrimental effects on proteins but is observed to produce a slightly elevated level of methionine oxidization over that usually observed in in‐gel tryptic digestion. Copyright © 2001 John Wiley & Sons, Ltd.
ISSN:0951-4198
1097-0231
DOI:10.1002/rcm.555