Arabidopsis plastoglobules 2D-gel separation

Protein complexes separation of isolated plastoglobules (PGs) from Arabidopsis thaliana (Col-0 ecotype). PGs were isolated from the plants when they were 4 weeks old under long day photoperiod ( during the morning in dark adapted plants,16 hours of light and 8 darkness photoperiod, LED light of 120 uE intensity). The PGs were solubilized with 1.1 % n-Dodecyl--D-Maltoside and separated in a Blue-Native gel. The gel lanes were excised and further separated in a denaturing second dimension by SDS-PAGE. The resulting gel was visualized by silver staining and the protein spots excised for in-gel trypsin digestion. Samples from the 2D gel were subjeted to shotgun proteomics. The protein samples were measured on an LTQ Orbitrap (Thermo Fisher Scientific Inc, Waltham Massachusetts, USA) coupled to a Rheos Allegro liquid chromatograph (Flux Instruments GmbH, Klaus Kaiser Hochstr. 48, 4002 Basel, CH).