Using iTRAQ® to Understand Cellular Processes and Environmental Effects on Proteins
Proteomics and mass spectrometry have provided unprecedented tools for fast, accurate, high throughput biomolecular separation and characterization, which are indispensable towards understanding biological and medical systems.
iTRAQ® technology uses different chemical tags that allow multiplexing of up to eight samples and produces identical MS/MS sequencing ions for all four versions of the same derivatized tryptic peptide.
Advantages of iTRAQ®:
- Avoid potential problems with 2D gel approach
- Wider diversity of proteins analyzed – hydrophobic, acidic and basic proteins
- Quantification at the peptide level
- iTRAQ® combined with multidimensional LC can analyze complex samples
- Up to eight samples can be analyzed simultaneously
The optimum sample amount is approximately 50-100 ?g per sample. The labeling is achieved at the peptide level. All the peptides are labeled. Quantitation is achieved by comparison of the peak areas and resultant peak ratios for the reporter ions in the MS/MS spectra, which reflect the relative abundance of the proteins.
Studying at the protein level allows researchers to investigate how proteins, their dynamics, modifications, and interactions affect cellular processes and how cellular processes and environment affect proteins.
If you have questions or would like to consult about using iTRAQ® for your research, please email the Mass Spectrometry core lab at ICBR-MassSpectrometry@ad.ufl.edu, or call 352-273-8030.