2000
DOI: 10.1002/1522-2683(200011)21:17<3666::aid-elps3666>3.0.co;2-6
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A modified silver staining protocol for visualization of proteins compatible with matrix-assisted laser desorption/ionization and electrospray ionization- mass spectrometry

Abstract: The growing availability of genomic sequence information, together with improvements in analytical methodology, have enabled high throughput, high sensitivity protein identification. Silver staining remains the most sensitive method for visualization of proteins separated by two-dimensional gel electrophoresis (2-D PAGE). Several silver staining protocols have been developed which offer improved compatibility with subsequent mass spectrometric analysis. We describe a modified silver staining method that is ava… Show more

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Cited by 661 publications
(295 citation statements)
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References 26 publications
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“…Spot plugs were destained and proteins were digested with trypsin as described previously. 42 All samples were run on a Finnigan LTQ mass spectrometer connected to a Surveyor chromatography system incorporating an auto-sampler. Tryptic peptides were resuspended in 0.1% formic acid and were separated by means of a modular CapaLC system (Finnigan) connected directly to the source of the LTQ.…”
Section: Discussionmentioning
confidence: 99%
See 1 more Smart Citation
“…Spot plugs were destained and proteins were digested with trypsin as described previously. 42 All samples were run on a Finnigan LTQ mass spectrometer connected to a Surveyor chromatography system incorporating an auto-sampler. Tryptic peptides were resuspended in 0.1% formic acid and were separated by means of a modular CapaLC system (Finnigan) connected directly to the source of the LTQ.…”
Section: Discussionmentioning
confidence: 99%
“…For preparative gels, the PlusOne Silver Staining kit (Amersham Biosciences, Uppsala, Sweden) was used with modifications to allow subsequent use with mass spectrometric analysis. 42 Image analysis Silver-stained gels were scanned at 100 mm resolution using a Personal SI laser densitometer (Molecular Dynamics, Sunnyvale, CA, USA). Protein spots were analysed quantitatively using the image analysis system Progenesis V2003.2 (Nonlinear Dynamics, Newcastle, UK).…”
Section: Samplesmentioning
confidence: 99%
“…The isoelectric focusing gel strips were loaded on top of 9% to 16% gradient polyacrylamide gels (200 3 250 3 1.0 mm) for a second round of SDS-PAGE using the Hoefer IsoDALT apparatus (Amersham-Pharmacia Biotech) until the tracking dye reached the anode ends of the gels. The gels were stained with silver nitrate according to the modified procedure by Yan et al (2000). Silver-stained gels were scanned using a high-resolution scanner (Bio-Rad GS-800 calibrated imaging densitometer).…”
Section: -De Analysis and Protein Identification By Maldi-tof Msmentioning
confidence: 99%
“…Gels underwent silver staining using a MALDI-TOF compatible protocol, according to Yan and co-authors (14). Two-dimensional electrophoresis 2DE was performed according to Gö rg and co-authors (15).…”
Section: Laboratory Methodsmentioning
confidence: 99%