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Fe-NTA Phosphopeptide Enrichment Kit

Fast and efficient phosphopeptide enrichment using Fe-NTA spin columns.

Pierce Fe-NTA Resin Kit for enrichement of phosphopeptides.

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).


  • 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.

Pierce Fe-NTA resin recovers more phosphorylated peptides than Sigma Phos-Select Reagent.
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.
  Thermo Scientific
Pierce Fe-NTA
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

Pierce Fe-NTA resin recovers multi-phosphorylated peptides.
The Thermo Scientific Pierce Fe-NTA Phosphopeptide Enrichment Kit effectively captures phosphopeptides containing multiple phosphates.

Related Resources:

Mass Spectrometry Sample Prep Workflow
Sample preparation for mass spectrometry technical guide

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

Ordering Information
Product # Description Pkg. Size Instructions MSDS CofA Price
88300 Fe-NTA Phosphopeptide Enrichment Kit
Formulation: Multi-component kit with Fe-NTA Spin Columns
Sufficient For: Processing 30 samples (200μL each) containing up to 150μg of phosphopeptides each

Kit Contents:
Fe-NTA Spin Columns, 30 columns
Binding Buffer, 6mL
Wash Buffer (2X), 12mL
Elution Buffer, 6mL
30-column kit Product Instructions for product #88300 Fe-NTA Phosphopeptide Enrichment Kit MSDS for product #88300 Fe-NTA Phosphopeptide Enrichment Kit Certificate of Analysis for product #88300 Fe-NTA Phosphopeptide Enrichment Kit $300.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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