Differences between iTRAQ® Reagents Application Kit - Protein (4374321) and iTRAQ® Reagent 4Plex or 8Plex Kits

For research use only. Not for use in diagnostic procedures.

Answer

The iTRAQ® Reagents Application Kit - Protein (PN 4374321) can be described as a protein-level approach where the protein of interest is labeled with iTRAQ reagent (either the 114, 115, 116 or 117 reagent depending on the protein state or other experimental conditions). The iTRAQ-labled protein undergoes electrophoresis using a SDS or native gel, followed by in-gel digestion and MS/MS analysis of labeled and unlabeled peptides.

Labeling proteins with iTRAQ reagents is an application that can be used by researchers to answer questions on protein topology or covalent complexes. Additional applications include relative quantification during the protein purification process.  

The more common mulitplexing iTRAQ reagents workflow follows a bottom-up approach, whereby many protein samples are denatured, alkylated and digested with trypsin. Peptides are then labelled with either the 4plex or 8plex iTRAQ reagents, as outlined in the workflows below.

Example of the iTRAQ reagent - 4 plex workflow: 

Example of the iTRAQ reagent - 8plex workflow:


Protocols for more details can be found here: https://sciex.com/products/consumables/itraq-reagent