Thermo Scientific Pierce Fe-NTA Phosphopeptide Enrichment Kit enables fast and efficient enrichment of phosphorylated peptides to process protein digests or strong cation-exchange peptide fractions for analysis by mass spectrometry.
The Pierce Fe-NTA Phosphopeptide Enrichment Kit enriches phosphopeptides from complex samples using iron-chelate resin in spin columns. These columns enrich a higher percentage of phosphopeptides than other resins and with an overall higher number of total and unique phosphopeptides. The complete kit is easy to use and requires less than one hour to process protein digests or strong cation-exchange peptide fractions for analysis by mass spectrometry (MS).
Highlights:
- Convenient – spin-column format enables parallel processing of multiple samples
- High capacity – each column enriches up to 150µg of phosphopeptides
- Excellent recovery – our kit enriches more total and unique phosphopeptides than other commercially available resins
- High coverage – recover peptides with single and multiple phosphorylation sites
Product Details:
To achieve robust MS results, enrichment of phosphopeptide samples is essential because of low abundance and poor ionization relative to non-phosphorylated peptides. Multiple phosphorylated amino acids within a peptide contribute to the complexity of phosphopeptide analysis. The Pierce Fe-NTA Spin Columns effectively enriches singly phosphorylated peptides as well as peptides with three or more phosphorylated sites, reducing sample complexity and enabling effective identification and characterization of phosphorylated peptides by MS.
The Pierce Fe-NTA Phosphopeptide Enrichment Kit contains detailed instructions and all necessary components to load, wash and elute phosphopeptides within an hour. This kit is compatible with samples digested in solution or after in-gel digestion. Using the Pierce Graphite Spin Columns (Part No. 88302) after enrichment will improve phosphopeptide analysis by efficiently binding hydrophilic peptides and efficiently removing urea, salts and other contaminants before MS analysis.
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| The Thermo Scientific Pierce Fe-NTA Phosphopeptide Enrichment Kit enriched a greater percentage of total and unique phosphopeptides from U2-OS cell lysate. Numbers refer to the total and unique number of phosphopeptides identified in each condition. |
| Average phosphopeptide enrichment results from duplicate experiments. Peptide summary results were analyzed with Thermo Scientific Proteome Discoverer 2.1 and Proteome Software Scaffold 3.0, then exported and summarized with Microsoft Excel* and Access* Programs. |
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Thermo Scientific
Pierce Fe-NTA |
Sigma
Phos-Select Reagent |
| Total phosphopeptides |
862 |
430 |
| Total peptides |
1753 |
1665 |
| Total unique peptides |
393 |
395 |
| Total unique phosphopeptides |
178 |
90 |
| Total phosphopeptides (%) |
53 |
31 |
| Unique phosphopeptides (%) |
50 |
27.5 |
 |
| The Thermo Scientific Pierce Fe-NTA Phosphopeptide Enrichment Kit effectively captures phosphopeptides containing multiple phosphates. |
Related Resources:
Mass Spec Sample Preparation Handbook
Mass Spectrometry Sample Prep Workflow
Review of sample preparation for mass spectrometry
Related Products:
TiO2 Phosphopeptide Enrichment and Clean-up Kit
Pierce Graphite Spin Columns
In-gel Tryptic Digestion Kit
Pierce Phosphoprotein Enrichment Kit
Pierce Magnetic Phosphopeptide Enrichment Kit
Halt Phosphatase Inhibitor Cocktails
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