METHODS
Nanoflow LC
Nanoflow liquid chromatography has become an invaluable tool and is the technique of choice in many proteomics applications. Restrictions on sample amounts and the essential demand for increased sensitivity can only be overcome by the use of dedicated nano LC systems. The UltiMate 3000 (Dionex) is a verstaile nano LC system operating routinely at flow rates of 300-350 nl/min. It is controlled by Chromeleon, an adaptable chromatography software. The tryptic peptides are separated on a peptide trap and a C18 column with a mobile phase formed from (A) HPLC grade water containing 2% acetonitrile and 0.1% formic acid and (B) HPLC grade water containing 90% acetonitrile and 0.1% formic acid. A nanospray source (Picoview) is interfaced to the mass spectrometers.
(click to enlarge)