Due to low stoichiometry of phosphorylation, phosphopeptide enrichment is a critical step for successful mass spectrometry-based phosphoproteomics experiments. PolyMAC offers an efficient and greatly improved method to achieve more complete phosphopeptide enrichment under homogeneous conditions. This highly selective enrichment procedure can be used with majority of the complex samples because it provides optimal specificity toward phosphopeptides PolyMAC-Ti M is a magnetic bead based purification method. Expedeon also offers PolyMAC-Ti G which is a spin column based method.
0.5mL of PolyMAC-Ti reagent
30mL of Loading buffer
20mL of Capture buffer
50mL of Washing buffer
20mL Elution buffer
In this method, a soluble nanopolymer is functionalized with metal ions to specifically capture O-phosphorylated peptides. PolyMAC-Ti offers an efficient and greatly improved method to achieve more complete phosphopeptide enrichment under homogeneous conditions.
100ug of digested complex cell lysate from DG-75 B-cell lymphoma cells was used to compare the enrichment efficiency between titanium-based PolyMAC and other commercially available sources of phosphopeptide enrichment reagents. A number of unique phosphosites identified and percent enrichment selectivity were used as the parameters for comparison. Each experiment was run in triplicate.