Microwave-assisted protein staining: mass spectrometry compatible methods for rapid protein visualisation

The effects of microwave irradiation on the staining of electrophoresed and electroblotted proteins have been assessed using currently available detection methods. Although the absorption of microwave radiation was found to be uneven, band intensity following microwave‐assisted protein staining (MAP...

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Bibliographic Details
Published in:Rapid communications in mass spectrometry 2002-01, Vol.16 (4), p.272-280
Main Authors: Nesatyy, Victor J., Dacanay, Andrew, Kelly, John F., Ross, Neil W.
Format: Article
Language:English
Subjects:
Databases, Factual
DNA Fingerprinting
Electrophoresis
Electrophoresis, Polyacrylamide Gel
Gold Colloid - analysis
Hydrolysis
Indicators and Reagents
Mass Spectrometry
Membranes, Artificial
Microwaves
Molecular Weight
Peptides - analysis
Polyvinyls
Proteins - chemistry
Rosaniline Dyes
Silver Staining
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Spectrophotometry, Ultraviolet
Trypsin - chemistry
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Summary:The effects of microwave irradiation on the staining of electrophoresed and electroblotted proteins have been assessed using currently available detection methods. Although the absorption of microwave radiation was found to be uneven, band intensity following microwave‐assisted protein staining (MAPS) was comparable and in some cases exceeded the intensity of the bands visualised by the original staining methods. It was found that microwave treatment drastically reduced the duration of the staining protocols for visualisation of the proteins separated by both one‐ and two‐dimensional electrophoresis. Application of MAPS methods did not affect peptide mass fingerprinting analysis by mass spectrometry and subsequent identification of the protein by database searching. The peptide mass maps corresponding to the proteins visualised using both the conventional and MAPS methods did not show significant difference in signal/noise ratio. Moreover, it appeared that microwave treatment of the gels resulted in the increased recovery of the peptides following in‐gel trypsin digestion. Briefly, microwave‐assisted protein staining methods were rapid, compatible with mass spectrometry and were equally effective on thin (0.75 mm) and thick (1.5 mm) gels (such as those used in 2D electrophoresis). Copyright © 2002 Crown in the right of Canada. Published by John Wiley & Sons, Ltd.
ISSN:0951-4198
1097-0231
DOI:10.1002/rcm.571