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BiochemistryPLoS ONE
Peer-reviewed journal article

Titration-WB: A methodology for accurate quantitative protein determination overcoming reproducibility errors

Alice MaestriiD, Ewa EhrenborgiD, Olivera Werngren, Maria Olin, Carolina E. HagbergiD, Matteo PedrelliiD, Paolo PariniiD

Abstract

Western blot (WB) technique is widely used to identify and quantify proteins in biological samples, but it is poorly accurate and precise for quantitative purposes. Merging the WB with the titration assay, we established a reliable method for protein quantification based on the slope of the linear function for signal-intensity/total protein loaded. Titration-WB unravels systematic and operator errors, compensates for problems in antibody affinity/avidity, and permits trustworthy inter-WB comparison.

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02 /Specification & Classification

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