Peptide Enrichment and Fractionation for Mass Spectrometry

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

	Fe-NTA Phosphopeptide Enrichment Kit	TiO2 Phosphopeptide Enrichment and Clean Up Kit	Magnetic TiO2 Phosphopeptide 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