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Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid
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Coomassie does it (better): A Robin Hood approach to total protein quantification
Nour Noaman1, Jens R Coorssen1
1Departments of Health Sciences and Biological Sciences, Faculties of Applied Health Sciences and Mathematics & Science, Brock University, St Catharines, Ontario, Canada.
Analytical Biochemistry
|May 16, 2018
Summary
A new, cost-effective method using colloidal Coomassie Brilliant Blue (cCBB) staining offers reproducible total protein quantification for proteomics. This adaptation of the EZQ™ assay provides accurate results for samples like human serum, improving quantitative proteomic analyses.
Area of Science:
- Proteomics
- Biochemistry
- Analytical Chemistry
Background:
- Accurate total protein quantification is crucial for reproducible quantitative proteomics.
- Established methods may have limitations in cost, reproducibility, or applicability to common buffers.
Purpose of the Study:
- To develop a straightforward, cost-effective adaptation of the EZQ™ total protein assay.
- To enhance reproducibility, sensitivity, and linearity for protein quantification in common solubilisation buffers.
Main Methods:
- Adaptation of the commercial EZQ™ protein quantification assay.
- Utilisation of non-proprietary materials and colloidal Coomassie Brilliant Blue (cCBB) staining.
- Testing with proteins in common solubilisation buffers and human serum.
Main Results:
- The adapted method demonstrated greater reproducibility and equal sensitivity compared to the original assay.
- Optimal signal linearity was achieved within a practical concentration range.
- More accurate and precise determinations of total protein concentration were obtained for human serum.
Conclusions:
- This adapted cCBB staining method offers a reliable and economical approach for total protein quantification.
- The method is well-suited for sample preparation in quantitative proteomic analyses, including those involving two-dimensional gel electrophoresis.
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