Coomassie does it (better): A Robin Hood approach to total protein quantification

Anal Biochem. 2018 Sep 1:556:53-56. doi: 10.1016/j.ab.2018.05.012. Epub 2018 Jun 28.

Abstract

Quantitative comparative proteomics require accurate and reproducible assessments of total protein concentration. We report a straightforward, cost-effective adaptation of an established commercial method for total protein quantification (EZQ™), utilising non-proprietary materials and colloidal Coomassie Brilliant Blue (cCBB) staining to achieve greater reproducibility, equal sensitivity, and optimal linearity of signal within a practical concentration range for proteins in common solubilisation buffers (i.e. for isoelectric focussing and/or SDS-PAGE). This method provided more accurate and precise determinations of total protein concentration in human serum prepared for two-dimensional gel electrophoresis, indicating it may be better suited as the lead-in to most quantitative proteomic analyses.

Keywords: Calibration standards; Dot-blot; EZQ™; Gel electrophoresis; Proteomics; Quantitation; Solid-phase assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Electrophoresis, Gel, Two-Dimensional*
  • Electrophoresis, Polyacrylamide Gel*
  • Proteins / analysis*
  • Proteomics
  • Rosaniline Dyes / chemistry*
  • Staining and Labeling*

Substances

  • Proteins
  • Rosaniline Dyes
  • coomassie Brilliant Blue