Fast Response Proteomics by Accelerated In-gel Digestion of Proteins

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Publikace nespadá pod Filozofickou fakultu, ale pod Přírodovědeckou fakultu. Oficiální stránka publikace je na webu muni.cz.
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HAVLIŠ Jan THOMAS Henrik ŠEBELA Marek SHEVCHENKO Andrej

Rok publikování 2003
Druh Článek v odborném periodiku
Časopis / Zdroj Analytical Chemistry
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
Obor Analytická chemie, separace
Klíčová slova in-gel digestion; quantitative analysis; mass spectrometry
Popis 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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