The DOI matches its Crossref record; 5 of the 6 references it cites were found in a registry or an index.
Protein quantification and its tolerance for different interfering reagents using the BCA-method with regard to 2D SDS PAGE
Abstract
Measuring the protein content of a sample is a mandatory and frequently practiced procedure in the lab. Although the procedure is quite simple and convenient to perform with commercially available kits, incompatible reagents in the lysate can cause problems in the quality of measurement. Unfortunately these reagents are cornerstones of high efficiency lysing buffers, e.g. high amounts of urea or beta-mercaptoethanol. In this study we addressed the tolerance of the well-known BCA-assay (bicinchoninic acid) to various reagents in different concentrations, with special regard to a subsequent 2D-gelelectrophoresis. As a result, the kit is incompatible with the recipes of regular 2D-buffers. Also, when mixing two different reagents interfering effects will occur in a non-predictable way. Therefore we established a new method to quantify protein content in lysates ready for 2D-gelelectrophoresis: by mixing an aliquot with SDS, an equilibration is performed to that the sample can be run on a regular 1D SDS PAGE. Image analysis following fluorescence staining (SYPRO Ruby) reveals the absolute protein content in comparison to a BSA dilution curve processed accordingly.
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“The methodology was validated considering the tolerance criteria described by Krieg et al. [16].”
From Functional and Bioactive Characterization of Hemp Cake Proteins and Polyphenols from Non-Psychoactive Cannabis sativa (2025) · cites this paper as Krieg et al. [16]
Citations
Cites
PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENT
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Bradford, M. M. (1976) · Analytical Biochemistry · doi:10.1016/0003-2697(76)90527-3 · Cited by 255,302
Measurement of protein using bicinchoninic acid
Smith, P. K., Krohn, R. I., Hermanson, G. T., Mallia, A. K., Gartner, F. H., Provenzano, M. D., Fujimoto, E. K., Goeke, N. M., Olson, B. J., & Klenk, D. C. (1985) · Analytical Biochemistry · doi:10.1016/0003-2697(85)90442-7 · Cited by 18,482
Leak, L. V., Liotta, L. A., Krutzsch, H. C., Jones, M., Fusaroa, V. A., Ross, S., Zhao, Y., & Petricoin, E. F. (2004) · PROTEOMICS · doi:10.1002/pmic.200300573 · Cited by 80
Dilution of protein gel stain results in retention of staining capacity
Krieg, R. C., Paweletz, C. P., Liotta, L. A., & Petricoin, E. F. (2003) · BioTechniques · doi:10.2144/03352pt01 · Cited by 16
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An overview of technical considerations for Western blotting applications to physiological research
Bass, J. J., Wilkinson, D. J., Rankin, D., Phillips, B. E., Szewczyk, N. J., Smith, K., & Atherton, P. J. (2016) · Scandinavian Journal of Medicine and Science in Sports · doi:10.1111/sms.12702 · Cited by 433
Erkekoğlu, P., Rachidi, W., Gürsoy-Yüzügüllü, Ö., Giray, B., Favier, A. E., Öztürk, M., & Hıncal, F. (2010) · Toxicology and Applied Pharmacology · doi:10.1016/j.taap.2010.07.016 · Cited by 207
Cortés‐Ríos, J., Zárate, A. M., Figueroa, J. D., Medina, J., Fuentes-Lemus, E., Rodriguez-Fernández, M., Aliaga, M. E., & López‐Alarcón, C. (2020) · Analytical Biochemistry · doi:10.1016/j.ab.2020.113904 · Cited by 181
Chitosan–hyaluronic acid hybrid film as a novel wound dressing: in vitro and in vivo studies
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Protein extraction from activated sludge: An analytical approach
Ras, M., Girbal‐Neuhauser, E., Paul, E., Spérandio, M., & Cavaillé-Lefebvre, D. (2007) · Water Research · doi:10.1016/j.watres.2007.11.011 · Cited by 154
Metadata
- Authors
- Krieg, R. C., Dong, Y., Schwamborn, K., & Knuechel, R.
- Affiliations
- RWTH Aachen University (DE)
- Venue
- Journal of Biochemical and Biophysical Methods
- Publisher
- Elsevier BV
- Published
- 2005-10
- Volume
- 65
- Issue
- 1
- Pages
- 13-19
- ISSN
- 0165-022X (ISSN-L)
- OpenAlex
- W2068769109
- Cited by
- 122
- References
- 6
- Citations by year
- 2026: 7 · 2025: 6 · 2024: 6 · 2023: 9 · 2022: 9 · 2021: 5 · 2020: 6 · 2019: 6 · 2018: 9 · 2017: 11 · 2016: 6 · 2015: 12 · 2014: 7 · 2013: 3 · 2012: 1
- FWCI
- 0.86
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Biochemical and Biophysical Methods (publishedVersion)
- PubMed (publishedVersion)
- RWTH Publications (RWTH Aachen) (submittedVersion)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced Biosensing Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Keywords
- protein quantification, bicinchoninic acid assay, urea, 2-mercaptoethanol
- MeSH terms
- Proteins, Quinolines, Reagent Kits, Diagnostic, Sensitivity and Specificity, Electrophoresis, Gel, Two-Dimensional, Reproducibility of Results
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