Fredens, Julius2; Engholm-Keller, Kasper2; Møller-Jensen, Jakob2; Larsen, Martin Røssel2; Færgeman, Nils J2
1 Department of Biochemistry and Molecular Biology, Faculty of Science, SDU2 Department of Biochemistry and Molecular Biology, Faculty of Science, SDU
Stable isotope labeling by amino acids combined with mass spectrometry is a widely used methodology for measuring relative changes in protein and phosphorylation levels at a global level. We have applied this method to the model organism Caenorhabditis elegans in combination with RNAi-mediated gene knockdown by feeding the nematode on pre-labeled lysine auxotroph Escherichia coli. In this chapter, we describe in details the generation of the E. coli strain, incorporation of heavy isotope-labeled lysine in C. elegans, and the procedure for a comprehensive global phosphoproteomic experiment.
Methods in Molecular Biology, 2014, Vol 1188, p. 107-24