ArticleAnalytical biochemistry2022
Stain-Free total-protein normalization enhances the reproducibility of Western blot data.
Article in Analytical biochemistry, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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28 citing papers in PubMed.
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- The Effect of Anti-Activin Receptor Type IIA and Type IIB Antibody on Muscle, Bone and Blood in Healthy and Osteosarcopenic Mice.Journal of cachexia, sarcopenia and muscle · 2025Article
- Titration-WB: A methodology for accurate quantitative protein determination overcoming reproducibility errors.PloS one · 2025Article
- Protocol for quantifying N-Myc and global protein translation in neuroblastoma cells using click chemistry on polyvinylidene fluoride membranes.STAR protocols · 2024Article
- Limiting 20S proteasome assembly leads to unbalanced nucleo-cytoplasmic distribution of 26S/30S proteasomes and chronic proteotoxicity.iScience · 2024Article
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5 authors.
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Abstract
We compared the accuracy of three common methods of total protein normalization. The Stain-Free method was accurate across different types/brands of western blotting membrane and for various protein loads, unlike Ponceau S and Amido Black. Normalizing to the housekeeping proteins Actin and β-Tubulin could match the accuracy of the Stain-Free method. However, compared to Actin or β-Tubulin, normalizing to the Stain-Free signal reduced variability that led to enhanced reproducibility and a reduction in the number of samples needed to obtain statistically significant results by >50%. Stain-Free normalization can enhance the reproducibility and hence the confidence in Western Blot data.
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