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

Warning

This publication doesn't include Faculty of Arts. It includes Faculty of Science. Official publication website can be found on muni.cz.

Authors	HAVLIŠ Jan SHEVCHENKO Andrej
Year of publication	2004
Type	Article in Periodical
Magazine / Source	Analytical Chemistry
MU Faculty or unit	Faculty of Science
Citation
Keywords	COMPARATIVE PROTEOMICS; PEPTIDE; MIXTURES; O-18; PHOSPHORYLATION; PURIFICATION; QUANTITATION; EXPRESSION; DIGESTION; STANDARDS
Description	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.