| Note |
The TMT labelled peptides were separated by immobilized pH gradient - isoelectric focusing (IPG-IEF) on a 3-10 strip. Peptides were extracted from the strips into 72 fractions by a prototype liquid handling robot, supplied by GE Healthcare Bio-Sciences AB. A plastic device with 72 wells was put onto each strip and 50 µl of ultra-pure water (Milli-Q, Millipore) was added to each well. After 30 minutes incubation, the liquid was transferred to a 96 well plate and the extraction was repeated 2 more times. The extracted peptides were dried in a SpeedVac vaccum concentrator and dissolved in 3% acetronitrile (ACN), 0.1 % formic acid. |