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.
02 /Specification & Classification
Click any tag to browse matching papers- Citations
- 4
- FWCI
- 0.96
- Field percentile
- top 29%
- Assay
- Research Focus
- Approach
Reproducibility ledger
Nobody has reported trying to reproduce this paper yet. Did you replicate an experiment or run the code? Your report saves the next lab a week.
Sign in to file a reproducibility report.
Related papers
- Validating Antibodies for Quantitative Western Blot Measurements with Microwestern Array
R. J. Koch, A. M. Barrette, … M. R. Birtwistle·bioRxiv·Nov 17, 2017
- MS Western, a method of multiplexed absolute protein quantification is a practical alternative to western blotting
M. Kumar, S. R. Joseph, … A. Shevchenko·bioRxiv·Oct 13, 2017
- Benchmarking accuracy and precision of intensity-based absolute quantification of protein abundances in Saccharomyces cerevisiae
Benjamín J. Sánchez, Petri‐Jaan Lahtvee, … Jens Nielsen·PROTEOMICSPeer-reviewed·Jan 16, 2021
- A Ponceau Staining based Dot-Blot Assay for reliable and cost-effective Protein Quantification
D.-L. Helbing, L. Böhm, … H. Morrison·bioRxiv·Sep 10, 2019
- FastCAT accelerates absolute quantification of proteins by using multiple short non-purified chimeric standards
I. Rzagalinski, A. Bogdanova, … A. Shevchenko·bioRxiv·Dec 28, 2021
- How the West(ern) Was Won: Solutions for Immunoblotting Large and Small Proteins
Paula Llabata, Pere Llinàs‐Arias·Gene & Protein in DiseasePeer-reviewed·Jun 30, 2023