Automated online sequential isotope labeling for protein quantitation applied to proteasome tissue-specific diversity.

ABSTRACT: Quantitation of protein abundance is a vital component in the proteomic analysis of biological systems, which can be achieved by differential stable isotopic labeling. To analyze tissue-derived samples, the isotopic labeling can be performed using chemical labeling of the peptides post-digestion. Standard chemical labeling procedures often require many manual sample handling steps, reducing the accuracy of measurements. Here, we describe a fully automated, online (in nanoLC columns), labeling procedure, which allows protein quantitation using differential isotopic dimethyl labeling of peptide N termini and lysine residues. We show that the method allows reliable quantitation over a wide dynamic range and can be used to quantify differential protein abundances in lysates and, more targeted,

SUBMITTER: Raijmakers R 

PROVIDER: S-EPMC2556017 | biostudies-literature | 2008 Sep

REPOSITORIES: biostudies-literature