Coomassie staining as loading control in Western blot analysis
Abstract
In Western blotting, immunodetection of housekeeping proteins is routinely performed to detect differences in electrophoresis loading. The present work describes a much faster and simpler protein staining method, which is compatible with ordinary blocking conditions. In addition, the method can be used after immunodetection with superior linearity compared to ordinary staining methods. After immunoblotting and staining, protein bands can be further identified using peptide mass fingerprinting.
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Total protein provides a stable reference measure that accounts for variation in protein loading and in the amount of protein transferred to the membrane, and can have superior linearity over a range of concentrations when compared to using a single reference protein.
“Total protein provides a stable reference measure that accounts for variation in protein loading and in the amount of protein transferred to the membrane (Welinder and Ekblad, 2011), and can have superior linearity over a range of concentrations when compared to using a single reference protein (Aldridge et al., 2008; Eaton et al., 2013).”
From Sex-specific developmental alterations in DYRK1A expression in the brain of a Down syndrome mouse model (2024) · cites this paper as Welinder and Ekblad, 2011
Total protein as a loading control can provide a more consistent measure for sample normalization than use of a single housekeeping protein, as it may show better linearity with variation in protein loading and controls for differences in protein transfer to the membrane during Western blot.
“The choice to use total protein as a loading control was informed by demonstrations that it can provide a more consistent measure for sample normalization than use of a single housekeeping protein, as it may show better linearity with variation in protein loading (Eaton et al., 2013) and it controls for differences in protein transfer to the membrane during the Western blot procedure (Welinder and”
From Sex-specific developmental alterations in DYRK1A expression in the brain of a Down syndrome mouse model (2024) · cites this paper as Welinder and Ekblad, 2011
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“…measure for sample normalization than use of a single housekeeping protein, as it may show better linearity with variation in protein loading (Eaton et al., 2013) and it controls for differences in protein transfer to the membrane during the Western blot procedure (Welinder and Ekblad, 2011).”
- Authors
- Welinder, C., & Ekblad, L.
- Affiliations
- Lund University (SE)
- Corresponding author
- Ekblad, Lars
- Venue
- Journal of Proteome Research
- Publisher
- American Chemical Society (ACS)
- Published
- 2011-02-03
- Volume
- 10
- Issue
- 3
- Pages
- 1416-1419
- ISSN
- 1535-3893 (ISSN-L)
- 1535-3907
- OpenAlex
- W1993988048
- Cited by
- 463
- References
- 0
- Citations by year
- 2026: 15 · 2025: 10 · 2024: 19 · 2023: 25 · 2022: 28 · 2021: 25 · 2020: 46 · 2019: 35 · 2018: 43 · 2017: 40 · 2016: 45 · 2015: 41 · 2014: 40 · 2013: 25 · 2012: 18
- FWCI
- 7.49
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Proteome Research (publishedVersion)
- PubMed (publishedVersion)
- Lund University Publications (Lund University) (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
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Keywords
- Coomassie staining, loading control, housekeeping proteins, protein staining, Western blot, peptide mass mapping
- MeSH terms
- Animals, Peptide Mapping, Proteins, Rosaniline Dyes, Staining and Labeling, Blotting, Western, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
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