Fast and efficient phosphopeptide enrichment using Fe-NTA spin columns
The Thermo Scientific Pierce Fe-NTA Phosphopeptide Enrichment Kit enables fast and efficient enrichment of phosphorylated peptides.
The complete kit is easy to use and requires less than 1 hour to process protein digests or strong cation-exchange peptide fractions for analysis by mass spectrometry (MS).
Highlights of the Pierce Fe-NTA Phosphopeptide Enrichment Kit:
- 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
About the Pierce Fe-NTA Phosphopeptide Enrichment Kit
To achieve robust MS results, enrichment of phosphopeptide samples is essential because of low abundance and poor ionization relative to non-phosphorylated peptides. 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.
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.
 |
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.† |
| |
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 |
| † Peptide summary results were exported from Scaffold and analyzed and summarized with Microsoft Excel* and Access*. |
 |
The Thermo Scientific Pierce Fe-NTA Phosphopeptide Enrichment Kit effectively captures phosphopeptides containing multiple phosphates. |
Related Resources:
Mass Spec Sample Handbook
Mass Spectrometry Sample Prep Workflow
Related Products:
Pierce Graphite Spin Columns
In-gel Tryptic Digestion Kit
Pierce Phosphoprotein Enrichment Kit
Pierce Magnetic Phosphopeptide Enrichment Kit
Halt Phosphatase Inhibitor Cocktails
*Trademark; see link in page footer
|