HAVLIŠ, Jan, Henrik THOMAS, Marek ŠEBELA and Andrej SHEVCHENKO. Fast Response Proteomics by Accelerated In-gel Digestion of Proteins. Analytical Chemistry. Washington, D.C., USA: American Chemical Society, 2003, vol. 75, No 6, p. 1300-1306, 6 pp. ISSN 0003-2700.
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Basic information
Original name Fast Response Proteomics by Accelerated In-gel Digestion of Proteins
Authors HAVLIŠ, Jan, Henrik THOMAS, Marek ŠEBELA and Andrej SHEVCHENKO.
Edition Analytical Chemistry, Washington, D.C., USA, American Chemical Society, 2003, 0003-2700.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10406 Analytical chemistry
Country of publisher United States of America
Confidentiality degree is not subject to a state or trade secret
Impact factor Impact factor: 5.250
Organization unit Faculty of Science
UT WoS 000181675900011
Keywords in English in-gel digestion; quantitative analysis; mass spectrometry
Tags in-gel digestion, mass spectrometry, Quantitative Analysis
Tags International impact, Reviewed
Changed by Changed by: doc. Mgr. Jan Havliš, Dr., učo 743. Changed: 2/7/2009 18:59.
Abstract
Kinetics of in-gel digestion of proteins by modified and native trypsins was studied by MALDI TOF mass spectrometry using 18O-labelled peptides as internal standards. The effect of the temperature, enzyme concentration, digestion time and surface area of gel pieces on the yield of digestion products was characterised. Based on the kinetic data we developed a protocol that enabled the identification of gel separated proteins with 30 min digestion time without compromising the peptide yield and the sensitivity compared to conventional protocols that typically rely upon overnight enzymatic cleavage. The accelerated digestion protocol was tested in identification of more than 120 proteins from budding and fission yeast on the low picomole level.
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