Thermo Scientific™ High Select™ Phosphopeptide Enrichment Kits & Reagents

The expanded High Select family of complete kits supports enrichment of phosphorylated peptides using Fe-NTA and TiO₂ immobilized resins in spin column, tip, and magnetic bead formats designed for use with MS analysis.

Product Code	Brand	Product Line	Description
15620306	Thermo Scientific™ A32992	High Select™	Fe-NTA Phosphopeptide Enrichment Kit
17727463	Thermo Scientific™ A52283	High Select™	Fe-NTA Magnetic Phosphopeptide Enrichment Kit
17737463	Thermo Scientific™ A52284	High Select™	Fe-NTA Magnetic Agarose
15693804	Thermo Scientific™ A32993	High Select™	TiO2 Phosphopeptide Enrichment Kit
10467734	Thermo Scientific™ 88302	Pierce™	Graphite Spin Columns
11812134	Thermo Scientific™ 88303	Pierce™	TiO2 Phosphopeptide Enrichment Tips

Description

Mass spectrometry (MS) is a key tool for identifying sites of protein phosphorylation and quantifying phosphorylation changes. However, MS analysis of protein phosphorylation is challenging due to the low stoichiometry, high hydrophilicity, poor ionization, and incomplete fragmentation of phosphopeptides. Given the low relative abundance of phosphorylation modifications in complex protein samples, enrichment is essential for successful MS analysis of phosphopeptides. High-Select Fe-NTA and TiO₂ phosphopeptide enrichment kits complement our lysis, reduction, alkylation, and digestion reagents, along with our C18, graphite spin, and high-pH reversed-phase peptide fractionation columns to provide a complete workflow for phosphopeptide enrichment.

• Convenient—pre-formulated buffers for ease-of-use in spin-column (Fe-NTA and TiO₂) or magnetic (Fe-NTA) formats to enable parallel processing of multiple samples
• High specificity—recovers phosphopeptides with > 90% selectivity
• Superior recovery—enriches more total and unique phosphopeptides than other commercially available resins
• High coverage—recovers peptides with single and multiple phosphorylation sites
• Scalable—compatible with automation, including the use of KingFisher magnetic particle processors

High-Select Fe-NTA Phosphopeptide Enrichment Kit enriches phosphopeptides from complex samples using iron-chelate resin in spin columns. Each column can enrich phosphopeptides from 0.5 to 5 mg of total protein digest as starting sample. Together with pre-formulated buffers, the improved and simplified procedure enriches up to 150 μg of phosphopeptides with greater than 90% selectivity in less than 45 minutes.

High-Select TiO2 Phosphopeptide Enrichment Kit includes 24 titanium dioxide (TiO2) spin tips with optimized buffers to facilitate preparation of enriched phosphopeptides. Each column can enrich phosphopeptides from 0.5 to 3 mg of total protein digest as starting sample. The optimized buffers and simplified procedure eliminate the need for pyrrolidine in the elution buffer and subsequent clean-up using graphite spin columns. Use of this kit results in enrichment of up to 150 μg of phosphopeptides with greater than 90% selectivity. TiO2 enriches a unique set of phosphopeptides that complements the High-Select Fe-NTA IMAC Phosphopeptide Enrichment Kit.

High-Select Fe-NTA Magnetic Phosphopeptide Enrichment Kit enriches phosphopeptides from complex samples using iron-chelate magnetic agarose beads. Each reaction is sufficient to enrich 250–1000 μg of phosphopeptides from 0.5–5 mg of total protein digest as starting sample in less than 45 minutes. Additionally, the kit may also be used to enrich for chemically modified peptides with functional groups designed for use with immobilized metal affinity chromatography resins.

High Select Fe-NTA Magnetic Agarose has been specifically designed to enrich phosphopeptides and chemically modified peptides for MS analysis. Our non-aggregating, magnetite (Fe₃O₄), superparamagnetic beads have exceptional uniformity for high performance using both manual and automated purification applications. The Fe-NTA Magnetic agarose is available separately to provide flexibility in experimental design.

For high resolution analysis of the enriched phosphopeptides, the recommended LC column for Nanospray Flex source is Acclaim PepMap 100 C18 LC Column (Cat. No. 164942 or 164939). For EASY-Spray source, the recommended LC column is EASY-Spray C18 LC Column (Cat. No. ES902 or ES903). The stand-alone High Select Fe-NTA Magnetic Agarose beads provide flexibility for scale and throughput and may be used in both manual and automated platforms, including KingFisher 96 and KingFisher Flex magnetic particle processors.

Safety and Handling

Product Identifier

• High Select Phosphopeptide Enrichment Kits & Reagents

Signal Word

• Danger

Hazard Category

• Acute toxicity Category 4
• Carcinogenicity Category 1A
• Serious eye damage/eye irritation Category 1
• Serious eye damage/eye irritation Category 2
• Flammable liquid Category 2
• Flammable liquid Category 3
• Skin corrosion/irritation Category 2

Hazard Statement

• H225-Highly flammable liquid and vapour.
• H226-Flammable liquid and vapour.
• H302-Harmful if swallowed.
• H312-Harmful in contact with skin.
• H315-Causes skin irritation.
• H318-Causes serious eye damage.
• H319-Causes serious eye irritation.
• H332-Harmful if inhaled.
• H350-May cause cancer.

Precautionary Statement

• P101-If medical advice is needed, have product container or label at hand.
• P102-Keep out of reach of children.
• P201-Obtain special instructions before use.
• P202-Do not handle until all safety precautions have been read and understood.
• P210-Keep away from heat, hot surfaces, sparks, open flames and other ignition sources. No smoking.
• P233-Keep container tightly closed.
• P240-Ground/bond container and receiving equipment.
• P241-Use explosion-proof electrical/ventilating/lighting/ equipment.
• P243-Take precautionary measures against static discharge.
• P260-Do not breathe dust/fume/gas/mist/vapours/spray.
• P261-Avoid breathing dust/fume/gas/mist/vapours/spray.
• P264-Wash hands thoroughly after handling.
• P270-Do not eat, drink or smoke when using this product.
• P280-Wear protective gloves/protective clothing/eye protection/face protection.
• P301+P310-IF SWALLOWED: Immediately call a POISON CENTER/doctor /
• P302+P352-IF ON SKIN: Wash with plenty of water/soap.
• P303+P361+P353-IF ON SKIN (or hair): Take off immediately all contaminated clothing. Rinse skin with water/ shower.
• P305+P351+P338-IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing.
• P308+P313-IF exposed or concerned: Get medical advice/attention.
• P332+P313-If skin irritation occurs: Get medical advice/attention.
• P362+P364-Take off contaminated clothing and wash it before reuse.
• P370+P378-In case of fire: Use dry sand, dry chemical, or alcohol-resistant foam to extinguish.
• P403+P235-Store in a well-ventilated place. Keep cool.
• P405-Store locked up.
• P501b-Dispose of contents/container in accordance with local/regional/national/international regulations.

Supplemental information

• MIXTURE LIST-Contains : ammonium hydroxide,acetonitrile, trifluoroacetic acid, lactic acid, titanium hydroxide, ethanol