Reference: Austin RJ, et al. (2012) IQcat: multiplexed protein quantification by isoelectric QconCAT. Proteomics 12(13):2078-83

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Abstract


Expression of isotopically labeled peptide standards as artificial concatamers (QconCATs) allows for the multiplex quantification of proteins in unlabeled samples by mass spectrometry. We have developed a generalizable QconCAT design strategy, which we term IQcat, wherein concatenated peptides are binned by pI to facilitate MS-sample enrichment by isoelectric focusing. Our method utilizes a rapid (∼2 weeks), inexpensive and scalable purification of arg/lys labeled IQcat standards in the Escherichia coli auxotroph AT713. With this pipeline, we assess the fidelity of IQcat-based absolute quantification for ten yeast proteins over a broad concentration range in a single information-rich isoelectric fraction. The technique is further employed for a quantitative study of androgen-dependent protein expression in cultured prostate cancer cells.

Reference Type
Journal Article | Research Support, Non-U.S. Gov't
Authors
Austin RJ, Chang DK, Holstein CA, Lee LW, Risler J, Wang JH, Adams L, Krusberski NB, Martin DB
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