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Thermo Scientific™ High-Select™ Fe-NTA Magnetic Phosphopeptide Enrichment Kit
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The Thermo Scientific™ High-Select™ Fe-NTA Magnetic Phosphopeptide Enrichment Kits use an advanced iron-chelate magnetic bead system designed for rapid and highly specific enrichment of phosphorylated peptides from complex biological samples. The beads coupled with our optimized, pre-formulated buffers achieve greater than 90% specificity for phosphopeptide enrichment while accommodating a broad range of starting material inputs (25–1,000 μg total protein digest) in a streamlined protocol requiring less than 45 minutes. The technology demonstrates significant analytical advantages over conventional methods, including reduced sample input requirements (50–100 μg versus 250–500 μg), enhanced pH compatibility, improved reagent safety profiles, and extended stability (12 months).
The high-throughput capability of our High-Select™ Fe-NTA Magnetic Phosphopeptide Enrichment Kits enable processing of 96 samples on a KingFisher Apex platform within 2 hours with only 30 minutes of hands-on time, delivering LC/MS-ready samples while reducing overall sample preparation time by 75% compared to traditional enrichment methods. The beads are compatible with tandem mass tag labeling approaches, supporting enrichment of both TMT™- and TMTpro™-labeled peptides. For optimal analytical performance, we recommend the EASY-Spray C18 LC columns (Cat. No. ES903) for high-resolution mass spectrometric analysis.
Features:
- High Specificity Performance: Achieve >90% selectivity for phosphopeptide recovery
- Superior Recovery Efficiency: Enrich more total and unique phosphopeptides than other commercially available resins
- Comprehensive Phosphorylation Coverage: Recover peptides with single and multiple phosphorylation sites
- Scalable Workflow Compatibility: Compatible with automated processing systems, including integration with KingFisher™ magnetic particle processors for high-throughput applications
Advanced Superparamagnetic Technology Ensures Superior Performance and Uniformity
The High-Select™ Fe-NTA Magnetic Phosphopeptide Enrichment Beads utilize cutting-edge non-aggregating, magnetite (Fe3O4) superparamagnetic technology that delivers exceptional uniformity across all applications. This advanced bead composition provides consistent and reliable performance in both manual purification protocols and automated systems, including compatibility with sophisticated instrumentation such as the Thermo Scientific™ KingFisher™ Apex Magnetic Particle Processor. The superparamagnetic properties of these magnetite-based beads ensure rapid magnetic separation while maintaining sample integrity throughout the enrichment process.
Streamlined Protocol Design Optimizes Phosphopeptide Recovery
The High-Select™ Fe-NTA Magnetic Phosphopeptide Enrichment Kits follow a systematic workflow that maximizes phosphopeptide capture efficiency. The protocol begins with standard peptide sample preparation involving lysis, reduction, alkylation, and digestion, followed by essential peptide cleanup and SpeedVac concentration steps. The magnetic beads must first be blocked to prevent non-specific binding interactions. Subsequently, peptide samples are reconstituted in binding/wash buffer and combined with the blocked beads for a 30-minute incubation period at room temperature with continuous agitation. The enrichment process concludes with magnetic collection of the beads, removal of unbound peptides, execution of multiple stringent wash steps, and final elution of the enriched phosphopeptides.
Critical Solution for Mass Spectrometry Challenges in Phosphoproteomics
Mass spectrometry (MS) represents the gold standard analytical technique for identifying protein phosphorylation sites and quantifying dynamic phosphorylation changes in biological systems. However, MS analysis of protein phosphorylation presents significant technical challenges that necessitate specialized enrichment strategies. Phosphopeptides exhibit characteristically low stoichiometry in complex protein mixtures, demonstrate high hydrophilicity that affects chromatographic behavior, suffer from poor ionization efficiency during MS analysis, and often display incomplete fragmentation patterns that complicate spectral interpretation. The inherently low relative abundance of phosphorylation modifications within complex protein samples makes enrichment an essential prerequisite for successful MS-based phosphopeptide analysis and reliable phosphoproteomics research outcomes.
Specifications
Specifications
| Product Type | Fe-NTA Phosphopeptide Enrichment |
| Product Line | High Select |
| Content And Storage | • High-Select Fe-NTA Magnetic Beads, 0.2 mL • Blocking Buffer, 1.0 mL • Binding/Wash Buffer B, 7.0 mL • Wash Buffer A, 3.0 mL • Phosphopeptide Elution Buffer, 2.5 mL |
| Shipping Condition | Wet Ice |
| Starting Material | Protease-digested Protein |
| Final Product Type | Peptides |
| Detection Method | Mass Spectrometry |
| Format | Kit |
| Workflow Step | Peptide Enrichment |
| Quantity | 20 Reactions |
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For Research Use Only.
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