Pierce™ TiO2 Phosphopeptide Enrichment and Clean-up Kit

Catalog number: 88301

Thermo Scientific™  Related applications: Protein Mass Spectrometry Analysis

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Description

Thermo Scientific Pierce TiO2 Phosphopeptide Enrichment and Clean-up Kit enables fast, selective enrichment of phosphorylated peptides for mass spectrometry using titanium dioxide spin tips, graphite spin columns and optimized buffers.

Features of the TiO2 Phosphopeptide Enrichment and Clean-up Kit:

Convenient—spin-column format of titanium dioxide and graphite columns enable parallel processing and clean up of multiple samples in less than 2 hours
High capacity—each column enriches up to 300 µg of phosphopeptides
Complementary—TiO2 enriches a unique set of phosphopeptides that complements the Pierce Fe-NTA IMAC Phosphopeptide Enrichment Kit
High selectivity—recover phosphopeptides with >90% selectivity

The complete kit includes 24 titanium dioxide (TiO2) spin tips and graphite spin columns with buffers to facilitate preparation of enriched and desalted phosphopeptides for analysis by mass spectrometry (MS).

The Pierce TiO2 Phosphopeptide Enrichment and Clean-up Kit protocol includes stringent washing conditions that increase the selectivity for phosphopeptides to >95%. Because some of the salts in these stringent washes may still be present in the eluted samples, the Pierce TiO2 Phosphopeptide Enrichment and Clean-up Kit includes graphite spin columns to desalt and concentrate enriched phosphopeptides, resulting in more successful phosphopeptide analysis results.

This kit is compatible with samples digested in solution or after in-gel digestion with trypsin or other MS grade proteases. Using the Pierce TiO2 Phosphopeptide Enrichment and Clean-up Kit and the Pierce Fe-NTA IMAC Phosphopeptide Enrichment Kits will enable complementary sets of phosphopeptides to be removed from complex samples.

About the TiO2 Resins:
Titanium dioxide enrichment of phosphopeptide samples is an essential tool for comprehensive MS analysis of phosphopeptides. The unique phosphopeptide binding properties of TiO2 resin results in different selectivity and preferential enrichment of multiply phosphorylated peptides compared to immobilized chelated metal ions (IMAC). TiO2 resin typically enriches more phosphopeptides than IMAC resins, and TiO2 preferentially enriches monophosphorylated peptides.

TiO2 resin typically binds phosphopeptides with a higher affinity than IMAC and the preferential enrichment of singly phosphorylated peptides with TiO2 is likely due to difficulty in eluting multiply phosphorylated peptides. To enrich a more comprehensive set of phosphopeptides, the flow-through and washes from IMAC resin can be acidified and applied to TiO2 resins and the resulting eluates pooled for analysis. In addition, the number of phosphopeptides recovered can be significantly increased by pre-fractionation of samples with strong cation exchange chromatography to reduce sample complexity prior to enrichment with TiO2 or IMAC processing.

Related Products
Pierce™ TiO2 Phosphopeptide Enrichment Spin Tips
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Product Size: 24 reactions
Final Product: Peptide
Workflow Step: Sample Enrichment
Detection Method: Mass Spectrometry
Starting Material: Protease-digested protein, Peptides
For Use With (Equipment): Mass Spectrometer, Microcentrifuge

Contents & storage

Sufficient For: Processing 24 digested protein samples (0.3-1 mg each) containing up to 100 µg of phosphopeptides each
• TiO2 Spin Tips, 24 tips
• Trifluoroacetic Acid, 1 mL
• 90% Lactic Acid, 2 mL
• Pyrrolidine, 200 µL
• Centrifuge Column Adaptors, 6 adaptors
• Pierce Graphite Spin Columns, 24 columns

Store kit at 4°C.

Documents

Manuals & protocols