ArticleProtein science : a publication of the Protein Society2025
Enhanced protein extraction and quantification protocol for microsamples: An ultra-sensitive workflow for low-volume, low-concentration total protein lysates.
Article in Protein science : a publication of the Protein Society, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
1 citing paper in PubMed.
- Enhanced protein extraction and quantification protocol for microsamples: An ultra-sensitive workflow for low-volume, low-concentration total protein lysates.Protein science : a publication of the Protein Society · 2025Article
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Authors and funding
7 authors.
Funding
Abstract
Accurate protein quantification at low concentrations and small volumes is critical for advancing small-scale research, such as microvasculature studies. However, existing microscale protocols often require ≥5 μL of sample or highly concentrated lysates, limiting their applicability in contexts with scarce material. To overcome these limitations, we developed the Nano-Extraction BCA-Optimized Workflow (NEBOW), a novel method requiring only 2 μL of sample and capable of detecting protein concentrations as low as 0.01 mg/mL. Optimized for the NanoDrop™ One UV-Vis Spectrophotometer, this workflow demonstrated significantly enhanced sensitivity and reproducibility compared to the standard BCA assay. Paired t tests (p < 0.01) and TOST equivalence testing (15% margin) confirmed key differences, with the NEBOW method producing steeper standard curves and more consistent results at low concentrations. Bland-Altman analysis showed that standard BCA tends to overestimate protein levels, while the NEBOW method maintained accuracy across a range of low-input samples. Western blot validation supported the improved performance of the new workflow. This approach offers a reliable, cost-effective solution for protein quantification when sample availability is limited, without sacrificing accuracy or sample integrity.
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Registered trials
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