Post digestion, peptide enrichment and/or fractionation is often necessary to detect peptides from low-abundant proteins and/or for reducing sample complexity for proteomic analysis.

There are several strategies that can be deployed to increase target protein identifications: utilize post-translational modification (PTM) specific affinity binding, such as phosphopeptide enrichment, or through fractionation, such as high pH reversed-phase chromatography which separates peptides by hydrophobicity.

The novel Thermo Scientific™ ActivX™ probes and enrichment kits enable the specific targeting of several enzyme classes, including kinases, GTPases, and serine hydrolases, which are covalently labeled in the active site at the protein level and enriched after digestion.

  • Comprehensive—reagents and kits for multiple peptide enrichment and/or fractionation strategies
  • Optimized—all kits are designed to maximize protein yield and increase target protein I.D.
  • Convenient—easy-to-use spin columns, tips or magnetic formats enable more rapid processing of peptide samples
  • Flexible—kits can be used with either cultured mammalian cells or tissues
  • Validated—all products have been fully tested and processed samples have been analyzed using Thermo Scientific™ Mass Spectrometers

Choose the right peptide enrichment or fractionation products for your applications

  Fe-NTA Phosphopeptide Enrichment Kit TiO2 Phosphopetide Enrichment and Clean Up Kit Magnetic TiO2 Phosphopetide Enrichment Kit High pH Reversed Phase Fractionation Kit Kinase Enrichment Kit with ATP Probe
Target Phosphopeptides Phosphopeptides Phosphopeptides All peptides ATP binding sites
Binding/labeling mechanism Metal chelate affinity to phosphate groups Metal oxide affinity to phosphate groups Metal oxide affinity to phosphate groups Hydrophobic interaction Biotinylated ATP analog to active site lysine
Enrichment strategy IMAC-Agarose resin

Spherical porous TiO2

TiO2 coated magnetic particles Hydrophobic polymer based resin Immobilized streptavidin agarose resin
Loading capacity/rxn* 0.5–5 mg 0.5–3 mg 100 µg 10–100 µg Lyophilized
Format Spin column Tip Magnetic bead Spin column Spin column
# of elution fractions 1 1 1 8 1
Processing time 45–60 min 45–60 min 15 min 30–60 min 6.5 hr + digestion
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*Loading capacity per reaction is based on a typical mammalian cultured cell protein digest sample

Other products for peptide enrichment & fractionation for mass spec

Featured data

The Thermo Scientific Pierce Fe-NTA Phosphopeptide Enrichment Kit effectively captures phosphopeptides with multiple phosphates.

The Thermo Scientific™ Pierce™ Fe-NTA Phosphopeptide Enrichment Kit effectively captures phosphopeptides with multiple phosphates.

Number of Phosphates per Peptide

1 2 3 4 5 6 Total
TiO2 492 103 8 4 0 1 608
Fe-NTA IMAC 234 34 216 3 1 0 488
Overlap 155 0 1 0 0 0 156

Selective enrichment of singly and multiply phosphorylated phosphopeptides with TiO2 and Fe-NTA IMAC. Average phosphopeptide enrichment results from duplicate experiments showing the number ofphosphopeptides containing one or more phosphate per peptide enriched using either method. Peptide spectrum summary results were exported from Proteome Software Scaffold 3.0 and analyzed with Microsoft™ Excel™ Software.