ArticlePloS one2025
Superior normalization using total protein for western blot analysis of human adipocytes.
Article in PloS one, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed.
- Malformin A1-mediated cytotoxicity in ovarian cancer cells occurs through pyroptosis and autophagy.FEBS open bio · 2026Article
- A Dose-Response Study on Human FGF21 to Inhibit Necrosis, Spectrin Breakdown, and to Increase Intracellular Cold Shock Proteins in Cultured Cortical Neurons Subjected to Oxygen-Glucose Deprivation Injury.Molecular neurobiology · 2026Article
- Time-restricted feeding counters brown adipose tissue whitening and inflammation in obese mice under chronic light exposure.Nutrition research and practice · 2026Article
- TMEM92 shields DDX3X from TTC3-mediated degradation to confer chemoresistance in triple-negative breast cancer.Clinical and translational medicine · 2026Article
- Aerobic glycolysis drives differentiation of unilocular adipocytes.Journal of lipid research · 2026Article
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Authors and funding
5 authors.
Funding
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Abstract
Western blotting has been a pivotal method for analyzing protein expression since the late 1970s, yet there is no established consensus on an optimal normalization strategy. In this study, primary mature human adipocytes were used to investigate the robustness of housekeeping proteins and total protein (TP) as normalization references for western blotting. TP exhibited the lowest variance among technical replicates compared to all investigated housekeeping proteins and was a superior normalization reference for the chosen protein-of-interest. TP also demonstrated the closest alignment with expected values when loaded as a protein gradient, highlighting the dynamic strength of TP as a normalization standard. Additionally, TP consistently demonstrated lower intra- and inter-individual variability in comparison to housekeeping proteins investigated across three metabolically similar individuals. In conclusion, TP normalization is the preferred method for reliable protein expression analysis in primary mature human adipocytes.
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