Optimization of SILAC labelling of DCsOther protocol
To generate a proteomic map of DCs (WP2) which includes quantitative information that can be used further to study signalling cascades (WP5) it is also necessary to optimize the SILAC labelling of DCs. This work was performed by Christian A. Luber (PhD student). First, we have set up a system for SILAC labelling of bone marrow derived murine dendritic cells (BM-DC). The principal method for generating BM-DC with GM-CSF was adapted from Lutz et al. (J Immunol Methods 1999, 223: 77-92). After some modifications of the protocol the FACS analysis of DC, which were generated in conditions described by Lutz et al. or in modified SILAC conditions did not show differences in the expression levels of the common DC markers CD11c, CD80, CD86. Analysis by mass spectrometry showed successful labelling.