One Step PolyMAC for Homogeneous Enrichment of Phosphopeptides Prior to Mass Spectrometry Analysis
PolyMAC is based on a metal ion functionalized soluble nanopolymer to chelate phosphopeptides in a homogeneous aqueous environment. Compared to current strategies based on solid phase micro and nanoparticles, PolyMAC demonstrates outstanding reproducibility, exceptional selectivity, fast chelation times, and high phosphopeptide recovery from complex mixtures.
Due to low stoichiometry of phosphorylation, phosphopeptide enrichment is a critical step for successful phosphoproteomics experiments. Available here is a novel chemical strategy termed Polymer based Metal ion Affinity Capture (PolyMAC) for the isolation of phosphopeptides with exceptionally high reproducibility, selectivity, and sensitivity.
Advantages of One step PolyMAC
- Higher reproducibility between experiments and labs
- Single step phosphopeptide and PolyMAC capture
- Improved phosphopeptide recovery
- Exceptional selectivity
- Outstanding reproducibility
- Fast chelation times.
The insolution binding of phosphopeptides to PolyMAC and the covalent capture onto solid-phase beads all occur in a single step. At this point, we offer capture based on magnetic beads only.
All kits include titanium based PolyMAC (PolyMAC-Ti) with magnetic capture beads, loading buffer, two washing buffers and elution buffer.
A magnetic separator rack is not included.
Insolution phosphopeptide enrichment using PolyMAC-Ti
100ug of digested complex cell lysate from DG-75 B-cell lymphoma cells was used to compare the enrichment efficiency between PolyMAC-Ti 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.