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

HAVLIŠ, Jan and 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, vol. 76, No 11, p. 3029-3036, 7 pp. ISSN 0003-2700.

Basic information

Original name

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

Name (in English)

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

Authors

HAVLIŠ, Jan and Andrej SHEVCHENKO

Edition

Analytical Chemistry, Washington, D.C., USA, American Chemical Society, 2004, 0003-2700

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Other information

Type of outcome

Článek v odborném periodiku

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 5.450

Organization unit

Faculty of Science

UT WoS

000221755200015

Keywords in English

COMPARATIVE PROTEOMICS; PEPTIDE; MIXTURES; O-18; PHOSPHORYLATION; PURIFICATION; QUANTITATION; EXPRESSION; DIGESTION; STANDARDS

Tags

COMPARATIVE PROTEOMICS, DIGESTION, Expression, MIXTURES, O-18, peptide, phosphorylation, purification, quantitation, standards

Tags

International impact, Reviewed

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Abstract

In English

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.