An in vivo labeling strategy with leucine-d3 for quantitative proteomics applied to the study of muscle cell differentiation

Shao En Ong, Akhilesh Pandey, Blagoy Blagoev, Irina Kratchmarova, Minerva Fernandez, Hanno Steen, Dan B. Kristensen, Matthias Mann

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

The mass spectrometric methods were used for the identification and quantitation of complex protein mixtures. The stable isotope labeling by amino acids (SILAC) method was used for the labeling by amino acids in cell culture for the in vivo incorporation of specific amino acids into all mammalian proteins. The mammalian cell lines were grown in media lacking a standard essential amino acid and the cultured cells specifically incorporated the stable isotope containing amino acids. The results show that the advantages of using stable isotopes to label proteins are, SILAC requires no peptide labeling steps and since the extent of incorporation is virtually 100% there are no differences in labeling efficiency between one sample and the other.

Original languageEnglish (US)
Title of host publicationProceedings 50th ASMS Conference on Mass Spectrometry and Allied Topics
Pages323-324
Number of pages2
StatePublished - 2002
Externally publishedYes
EventProceedings - 50th ASMS Conference on Mass Spectrometry and Allied Topics - Orlando, FL, United States
Duration: Jun 2 2002Jun 6 2002

Other

OtherProceedings - 50th ASMS Conference on Mass Spectrometry and Allied Topics
CountryUnited States
CityOrlando, FL
Period6/2/026/6/02

ASJC Scopus subject areas

  • Spectroscopy

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    Ong, S. E., Pandey, A., Blagoev, B., Kratchmarova, I., Fernandez, M., Steen, H., Kristensen, D. B., & Mann, M. (2002). An in vivo labeling strategy with leucine-d3 for quantitative proteomics applied to the study of muscle cell differentiation. In Proceedings 50th ASMS Conference on Mass Spectrometry and Allied Topics (pp. 323-324)