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Related Concept Videos

Effects of EDTA on End-Point Detection Methods01:18

Effects of EDTA on End-Point Detection Methods

Different methods, such as visual observance of metal-ion indicators, spectroscopic techniques, and potentiometric methods, can determine the endpoint of an EDTA titration.
In the visual method, metal-ion indicators (metallochromic dyes), which have distinct colors in their free and complex forms, are added to the mixture to signal the titration's end point. They form stable complexes with metal ions, but these complexes are weaker than the corresponding metal–EDTA complexes. As a result, EDTA...

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Related Experiment Video

Updated: Jul 14, 2026

A Fast Silver Staining Protocol Enabling Simple and Efficient Detection of SSR Markers using a Non-denaturing Polyacrylamide Gel
10:27

A Fast Silver Staining Protocol Enabling Simple and Efficient Detection of SSR Markers using a Non-denaturing Polyacrylamide Gel

Published on: April 20, 2018

Silver- and Coomassie-staining protocols: detection limits and compatibility with ESI MS.

Christiane Winkler1, Katrin Denker, Stefanie Wortelkamp

  • 1Rudolf-Virchow-Center, DFG-Research Center for Experimental Biomedicine, University of Würzburg, Würzburg, Germany.

Electrophoresis
|May 23, 2007
PubMed
Summary

Two silver-staining protocols are recommended for mass spectrometry (MS) analysis after polyacrylamide gel electrophoresis (PAGE). These methods offer high sensitivity and sequence coverage, crucial for effective MS analysis of proteins.

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Fast and Sensitive Colloidal Coomassie G-250 Staining for Proteins in Polyacrylamide Gels
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Fast and Sensitive Colloidal Coomassie G-250 Staining for Proteins in Polyacrylamide Gels

Published on: August 3, 2009

Staining Proteins in Gels
10:55

Staining Proteins in Gels

Published on: July 8, 2008

Related Experiment Videos

Last Updated: Jul 14, 2026

A Fast Silver Staining Protocol Enabling Simple and Efficient Detection of SSR Markers using a Non-denaturing Polyacrylamide Gel
10:27

A Fast Silver Staining Protocol Enabling Simple and Efficient Detection of SSR Markers using a Non-denaturing Polyacrylamide Gel

Published on: April 20, 2018

Fast and Sensitive Colloidal Coomassie G-250 Staining for Proteins in Polyacrylamide Gels
10:57

Fast and Sensitive Colloidal Coomassie G-250 Staining for Proteins in Polyacrylamide Gels

Published on: August 3, 2009

Staining Proteins in Gels
10:55

Staining Proteins in Gels

Published on: July 8, 2008

Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Staining protocols for polyacrylamide gel electrophoresis (PAGE) must be sensitive.
  • Staining methods should not interfere with subsequent mass spectrometry (MS) analysis.
  • Optimizing staining for nano-liquid chromatography-tandem mass spectrometry (nano-LC-MS/MS) is critical for proteomic studies.

Purpose of the Study:

  • To systematically evaluate the compatibility of various silver- and Coomassie-staining protocols with nano-LC-MS/MS.
  • To compare the sensitivity and sequence coverage achieved by different staining methods.
  • To identify optimal staining protocols for protein analysis using nano-LC-MS/MS.

Main Methods:

  • Tested 13 silver-staining, 1 imidazole-staining, and 2 Coomassie-staining protocols.
  • Utilized three model proteins (bovine serum albumin, rabbit L-lactate dehydrogenase, bovine milk beta-lactoglobulin) at concentrations ranging from 12 pmol to 30 fmol.
  • Assessed sequence coverage and detection sensitivity for each protocol in conjunction with a nano-LC-MS/MS system.

Main Results:

  • Compared the performance of multiple staining protocols for PAGE.
  • Determined the impact of different staining methods on protein detection sensitivity and MS analysis.
  • Identified two specific silver-staining protocols that provide a balance of high sequence coverage and sensitivity.

Conclusions:

  • Two silver-staining protocols (Blum et al., 1987 and Shevchenko et al., 1996) are highly suitable for nano-LC-MS/MS analysis.
  • These recommended protocols effectively combine good sequence coverage with excellent detection sensitivity.
  • The findings provide practical guidance for researchers performing proteomic analyses requiring sensitive protein identification post-PAGE.