iTRAQ technology coupled with 1D or 2D LC-MS/MS for multiplexing 4-8 samples simultaneously in a single experiment is available. iTRAQ enables relative and absolute quantitation of proteins and peptides by labelling samples with isotope encoded reporter ions. Differential expression of proteins of interest can be determined.
The protein sample is acetone precipitated, digested and labelled. Four (or eight) different samples can be labelled with iTRAQ reagents. Labelled peptides are first separated by High pH reverse phase chromatography with fraction concatenation. Each concatenated fraction is further analysed by LC-MS/MS. Results are compared using a specialised software.
