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

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

Analytical chemistry (2003-11-01)
Ying-Qing Yu, Martin Gilar, Peter J Lee, Edouard S P Bouvier, John C Gebler
초록

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.

MATERIALS
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Sigma-Aldrich
Nα-Benzoyl-L-arginine ethyl ester hydrochloride, trypsin substrate