Protein Quantification in Drosophila Heads

Sample preparation, western-blotting and quantification were performed as published by Trautenberg et al. (2019) (link). In brief, flies were raised on normal food and 5 days-old adults were transferred onto respective experimental foods for 7 days at 20°C. Thereafter, flies were snap-frozen in liquid nitrogen and frozen heads from females collected for processing. Heads were homogenized on ice in 0.01% Tx100-PBS and subsequently cooked at 95°C for 5 min. Polyclonal antibodies used to probe were Akt-pSer505 (Cell Signaling, 4054S), Akt-pThr308 (Invitrogen, 44-602G), Akt (Invitrogen, MAS14916).