Fast Response Proteomics by Accelerated In-gel Digestion of Proteins

HAVLIŠ, Jan, Henrik THOMAS, Marek ŠEBELA a Andrej SHEVCHENKO. Fast Response Proteomics by Accelerated In-gel Digestion of Proteins. Analytical Chemistry. Washington, D.C., USA: American Chemical Society, 2003, roč. 75, č. 6, s. 1300-1306, 6 s. ISSN 0003-2700.

Základní údaje

Originální název

Fast Response Proteomics by Accelerated In-gel Digestion of Proteins

Autoři

HAVLIŠ, Jan, Henrik THOMAS, Marek ŠEBELA a Andrej SHEVCHENKO

Vydání

Analytical Chemistry, Washington, D.C., USA, American Chemical Society, 2003, 0003-2700

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Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10406 Analytical chemistry

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 5.250

Organizační jednotka

Přírodovědecká fakulta

UT WoS

000181675900011

Klíčová slova anglicky

in-gel digestion; quantitative analysis; mass spectrometry

Štítky

in-gel digestion, mass spectrometry, Quantitative Analysis

Příznaky

Mezinárodní význam, Recenzováno

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Anotace

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.