ArticleScientific reports2022
A critical path to producing high quality, reproducible data from quantitative western blot experiments.
Article in Scientific reports, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Who cites it
30 citing papers in PubMed.
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- Inhibition of protein or glutamine biosynthesis affect the light-induced dephosphorylation of the SBiP1 chaperone in Symbiodiniaceae.Bioscience reports · 2025Article
- A novel L-shaped ortho-quinone analog targeting adenosine A2b receptor to inhibit epithelial-mesenchymal transition in colorectal cancer cells.Medical oncology (Northwood, London, England) · 2025Article
- Defining Biological Variability, Analytical Precision and Quantitative Biophysiochemical Characterization of Human Urinary Extracellular Vesicles.Journal of extracellular vesicles · 2025Article
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- Screening of Plant UDP-Glycosyltransferases for Betanin Production in Yeast.Applied biochemistry and biotechnology · 2025Article
- Fluorescence-Based Multiplex Western Blot to Simultaneously Detect the Insulin-Like Growth Factor-1 (IGF-1) Isoforms.Electrophoresis · 2025Article
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4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Western blotting experiments were initially performed to detect a target protein in a complex biological sample and more recently, to measure relative protein abundance. Chemiluminescence coupled with film-based detection was traditionally the gold standard for western blotting but accurate and reproducible quantification has been a major challenge from this methodology. The development of sensitive, camera-based detection technologies coupled with an updated technical approach permits the production of reproducible, quantitative data. Fluorescence reagent and detection solutions are the latest innovation in western blotting but there remains questions and debate concerning their relative sensitivity and dynamic range versus chemiluminescence. A methodology to optimize and produce excellent, quantitative western blot results with rigorous data analysis from membranes probed with both fluorescent and chemiluminescent antibodies is described. The data reveal when and how to apply these detection methods to achieve reproducible data with a stepwise approach to data processing for quantitative analysis.
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