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Anal Chem. 2003 Nov 1;75(21):6023-8.

Enzyme-friendly, mass spectrometry-compatible surfactant for in-solution enzymatic digestion of proteins.

Author information

1
Life Sciences Research and Development, Waters Corporation, 34 Maple Street, Milford, Massachusetts 01757, USA.

Abstract

Improved in-solution tryptic digestion of proteins in terms of speed and peptide coverage was achieved with the aid of a novel acid-labile anionic surfactant (ALS). Unlike SDS, ALS solubilizes proteins without inhibiting trypsin or other common endopeptidases activity. Trypsin activity was evaluated in the presence of various denaturants; little or no decrease in proteolytic activity was observed in 0.1-1% ALS solutions (w/v). Sample preparation prior to mass spectrometry and liquid chromatography analysis consists of sample acidification. ALS degrades rapidly at low-pH conditions, which eliminates surfactant-caused interference with analysis. Described methodology combines the advantages of protein solubilization, rapid digestion, high peptide coverages, and easy sample preparation for mass spectrometry and liquid chromatography analyses.

PMID:
14588046
DOI:
10.1021/ac0346196
[Indexed for MEDLINE]

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