MS Western, a Method of Multiplexed Absolute Protein Quantification is a Practical Alternative to Western Blotting.

ABSTRACT: Absolute quantification of proteins elucidates the molecular composition, regulation and dynamics of multiprotein assemblies and networks. Here we report on a method termed MS Western that accurately determines the molar abundance of dozens of user-selected proteins at the subfemtomole level in whole cell or tissue lysates without metabolic or chemical labeling and without using specific antibodies. MS Western relies on GeLC-MS/MS and quantifies proteins by in-gel codigestion with an isotopically labeled QconCAT protein chimera composed of concatenated proteotypic peptides. It requires no purification of the chimera and relates the molar abundance of all proteotypic peptides to a single reference protein. In comparative experiments, MS Western outperformed immunofluorescence Western

SUBMITTER: Kumar M 

PROVIDER: S-EPMC5795399 | biostudies-literature | 2018 Feb

REPOSITORIES: biostudies-literature