Fast Response Proteomics by Accelerated In-gel Digestion of Proteins

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.

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

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Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10406 Analytical chemistry

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

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

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Abstract

V originále

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.