On-line, Chip-Based Strategy for Increased Depth of Coverage in Proteomic Samples
On-line, Chip-Based Strategy for Increased Depth of Coverage in Proteomic Samples
8 Oct 2015On-line two-dimensional (2D) liquid chromatography is widely used for protein identification and quantification because of the advantage of increased peak capacity. A common workflow is the combination of strong cation exchange (SCX) as an orthogonal first dimension to reversed-phase (RP) chromatography as the second dimension. A more recently developed strategy has emerged where the first dimension is a high pH RP separation which has less orthogonality, but provides higher peak capacity compared to SCX. These 2D workflows are essential for measuring lower abundant proteins in complex proteomes, however can be complex to implement. In this application note, a simplified chip based 2D-LCMS workflow using a high pH/RP first dimensional separation and a low pH/RP secondary dimension coupled directly to mass spectrometer for proteomic analysis is described.
Related products
Request Quote for All Products
ProteinPilot software
SCIEXProteinPilot software is a powerful, robust and intuitive software tool for protein identification and quantification for discovery research and protein characterization. A paradigm shift in protein identification and relative protein expression analysis for protein research, ProteinPilot software streamlines protein identification and quantitation, enabling you to identify hundreds of peptide modifications and non-tryptic cleavages simultaneously. Powered by sophisticated algorithms, you can now easily and confidently distinguish protein isoforms, protein subsets, and suppress false positives, as well as visualize peptide-protein associations and relationships.