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 KitTiO2 Phosphopetide Enrichment and Clean Up KitMagnetic TiO2 Phosphopetide Enrichment KitHigh pH Reversed Phase Fractionation KitKinase Enrichment Kit with ATP Probe
TargetPhosphopeptidesPhosphopeptidesPhosphopeptidesAll peptidesATP binding sites
Binding/labeling mechanismMetal chelate affinity to phosphate groupsMetal oxide affinity to phosphate groupsMetal oxide affinity to phosphate groupsHydrophobic interactionBiotinylated ATP analog to active site lysine
Enrichment strategyIMAC-Agarose resin

Spherical porous TiO2

TiO2 coated magnetic particlesHydrophobic polymer based resinImmobilized streptavidin agarose resin
Loading capacity/rxn*0.5–5 mg0.5–3 mg100 µg10–100 µgLyophilized
FormatSpin columnTipMagnetic beadSpin columnSpin column
# of elution fractions11181
Processing time45–60 min45–60 min15 min30–60 min6.5 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

123456Total
TiO24921038401608
Fe-NTA IMAC23434216310488
Overlap15501000156

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.

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

123456Total
TiO24921038401608
Fe-NTA IMAC23434216310488
Overlap15501000156

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.