Archive: MS Western: The Method for Targeted Multiplexed Absolute Quantification of Proteins by GeLC-MS/MS

Absolute (in moles) quantification of proteins is required for the biomarker discovery in clinical proteomics, for modelling in system biology, to study protein kinetics, to determine the stoichiometry of the subunits in a multi-protein complex or to determine stoichiometric ratios between proteins and also non-proteinous ligands, such as lipids or DNA. However, currently available methods are technically demanding, expensive and often biased. For instance, accurate quantification by Western blotting (WB) requires highly specific monoclonal antibodies and pure protein standards. 

We have developed a method termed MS Western that accurately (CV <10%) determines the molar content of dozens of user-selected proteins at the low to sub-femtomole level in whole cell or tissue lysates without metabolic or chemical labelling or using specific antibodies.

Original Publication

Mukesh Kumar, Shai R Joseph, Martina Augsburg, Aliona Bogdanova, David Drechsel, Nadine L Vastenhouw, Frank Buchholz, Marc Gentzel and Andrej Shevchenko. MS Western, a method of multiplexed absolute protein quantification is a practical alternative to Western blotting. MCP (2017)
doi: 10.1074/mcp.O117.067082

Bharath Kumar Raghuraman, Sarita Hebbar, Mukesh Kumar, HongKee Moon, Ian Henry, Elisabeth Knust, Andrej Shevchenko. Absolute Quantification of Proteins in the Eye of Drosophila melanogaster. Proteomics, 20(23) Art. No. e1900049 (2020) DOI 


Kurzchalia lab
Knust lab

Dr. Bharath Kumar Raghuraman

Contact: raghuram(at)