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Anal Chem. 2013 Jan 15;85(2):907-14. doi: 10.1021/ac302423t. Epub 2013 Jan 7.

Mass spectrometry compatible surfactant for optimized in-gel protein digestion.

Author information

1
Promega Corporation, 2800 Woods Hollow Road, Madison, Wisconsin 53711, USA. sergei.saveliev@promega.com

Abstract

Identification of proteins resolved by SDS-PAGE depends on robust in-gel protein digestion and efficient peptide extraction, requirements that are often difficult to achieve. A lengthy and laborious procedure is an additional challenge of protein identification in gel. We show here that with the use of the mass spectrometry compatible surfactant sodium 3-((1-(furan-2-yl)undecyloxy)carbonylamino)propane-1-sulfonate, the challenges of in-gel protein digestion are effectively addressed. Peptide quantitation based on stable isotope labeling showed that the surfactant induced 1.5-2 fold increase in peptide recovery. Consequently, protein sequence coverage was increased by 20-30%, on average, and the number of identified proteins saw a substantial boost. The surfactant also accelerated the digestion process. Maximal in-gel digestion was achieved in as little as one hour, depending on incubation temperature, and peptides were readily recovered from gel eliminating the need for postdigestion extraction. This study shows that the surfactant provides an efficient means of improving protein identification in gel and streamlining the in-gel digestion procedure requiring no extra handling steps or special equipment.

PMID:
23256507
DOI:
10.1021/ac302423t
[Indexed for MEDLINE]

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