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

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

Author information

1
Departments of Health Sciences and Biological Sciences, Faculties of Applied Health Sciences and Mathematics & Science, Brock University, St Catharines, Ontario, Canada. Electronic address: n.noaman@westernsydney.edu.au.
2
Departments of Health Sciences and Biological Sciences, Faculties of Applied Health Sciences and Mathematics & Science, Brock University, St Catharines, Ontario, Canada. Electronic address: jcoorssen@brocku.ca.

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

PMID:
29763592
DOI:
10.1016/j.ab.2018.05.012

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