Fast Response Proteomics by Accelerated In-gel Digestion of Proteins

Varování

Publikace nespadá pod Filozofickou fakultu, ale pod Přírodovědeckou fakultu. Oficiální stránka publikace je na webu muni.cz.

Autoři	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.