Authors
Joern Dengjel, Vyacheslav Akimov, Blagoy Blagoev, and Jens Andersen
Summary
It has been shown that signaling via phosphorylation happens within seconds after growth factor stimulation in eukaryotic cells. However, these very early signaling events have so far not been analyzed by mass spectrometry (MS)-based proteomics. Here we report the development of an automated system which allows quantitative proteomic assessment of very early cellular signaling events (qPACE). Our approach relies on pumping SILAC labeled cells through a continuous quench-flow system to ensure rapid and reproducible initiation and trapping of cellular signaling states. The method permits accurate quantitative measurements by MS of signaling events, like phosphorylation, on a second timescale.Further details
The protocol was published on Protocol Exchange in 2007. To see the entire protocol, click on the source link.Advertisement
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