Thermo Scientific™ High Select™ Phosphopeptide Enrichment Kits & Reagents

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 TiO 2 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 TiO 2 ) 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 (Fe 3 O 4 ), 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.