Construction, growth, and harvesting of fission yeast stable isotope labeling by amino acids in cell culture (SILAC) strains

André Koch, Claudia C. Bicho*, Weronika E. Borek, Alejandro Carpy, Boris Macěk, Silke Hauf, Kenneth E. Sawin

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Stable isotope labeling by amino acids in cell culture (SILAC) enables the relative quantification of protein amounts and posttranslational modifications in complex biological samples through the use of stable heavy isotope–labeled amino acids. Here we describe methods for the application of SILAC to fission yeast Schizosaccharomyces pombe using either labeled lysine or a combination of labeled lysine and labeled arginine. The latter approach is more complicated than the use of labeled lysine alone but may yield a more comprehensive (phospho)proteomic analysis. The protocol includes methods for construction of SILAC-compatible strains, growth of cultures in labeled medium, cell harvesting, and protein extraction.

Original languageEnglish
Pages (from-to)463-471
Number of pages9
JournalCold Spring Harbor protocols
Volume2017
Issue number6
DOIs
Publication statusPublished - 1 Jun 2017

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