Absolute Quantification of Proteins in Solutions and in Polyacrylamide Gels by Mass Spectrometry

HAVLIŠ, Jan a Andrej SHEVCHENKO. Absolute Quantification of Proteins in Solutions and in Polyacrylamide Gels by Mass Spectrometry. Analytical Chemistry. Washington, D.C., USA: American Chemical Society, 2004, roč. 76, č. 11, s. 3029-3036, 7 s. ISSN 0003-2700.

Základní údaje

• Originální název: Absolute Quantification of Proteins in Solutions and in Polyacrylamide Gels by Mass Spectrometry
• Název anglicky: Absolute Quantification of Proteins in Solutions and in Polyacrylamide Gels by Mass Spectrometry
• Autoři: HAVLIŠ, Jan a Andrej SHEVCHENKO.
• Vydání: Analytical Chemistry, Washington, D.C., USA, American Chemical Society, 2004, 0003-2700.

Další údaje

• Typ výsledku: Článek v odborném periodiku
• Utajení: není předmětem státního či obchodního tajemství
• Impakt faktor: Impact factor: 5.450
• Organizační jednotka: Přírodovědecká fakulta
• UT WoS: 000221755200015
• Klíčová slova anglicky: COMPARATIVE PROTEOMICS; PEPTIDE; MIXTURES; O-18; PHOSPHORYLATION; PURIFICATION; QUANTITATION; EXPRESSION; DIGESTION; STANDARDS
• Štítky: COMPARATIVE PROTEOMICS, DIGESTION, Expression, MIXTURES, O-18, peptide, phosphorylation, purification, quantitation, standards
• Příznaky: Mezinárodní význam, Recenzováno

Anotace anglicky

A combination of nanoelectrospray tandem mass spectrometry and O-18-labeled peptide internal standards was applied for the absolute quantification of proteins from their in-solution and in-gel tryptic digests. Although absolute quantification from in-solution digests was accurate, we observed that in-gel digestion compromised the quantification accuracy by affecting the recovery of individual peptides and, therefore, the provided estimates might be strongly influenced by the selection of reference peptides. Under optimized experimental conditions, it was possible to provide a semiquantitative estimate of the absolute amount of gel separated proteins within better than 50% error margin.