Protein Phosphorylation

Phosphopeptide Enrichment Kit


Fast, easy, unbiased phosphopeptide capture for small samples.
Proteomics and bio-pathway researchers studying the dynamic cycles of phosphorylation demanded an enabling tool. We responded by making a complete kit for phosphopeptide generation and enrichment that handles the precious small samples you really use. Outputs are compatible with Liquid Chromatography and Mass Spectrometry. Use this tool in combination with powerful quantitative mass spectrometry methods like AQUA!

PhosphoProfile™ I Phosphopeptide Enrichment Kit

The complete solution kit – 24 samples up to 25 nmoles phosphophopeptide
  • Proteomics Grade Trypsin - for clean and complete digests
  • PHOS-Select™ Gallium Spin Columns – high capacity, novel Ga+3 silica media for fast,
    unbiased capture and recovery
  • Controls – validate your process for confidence and reporting
  • Buffers – enzyme reaction, binding, washing and elution optimized formulations for robust
  • Collection tubes – matched equipment means no additional purchases or waste
Workflow highlighting the use of the spin columns for detection and selective enrichment of phosphopeptides from sample.

HPLC Chromatograms comparing phosphopeptide enrichment by IMAC methods

HPLC Chromatograms comparing phosphopeptide enrichment by IMAC methods
Quantitation values are given in the table below. Note that competitor A, B, and C technologies were biased in selecting Peptide 2, while Sigma’s PHOS-Select™ Gallium technology efficiently bound and eluted the peptides in essentially the same ratio as applied to the column.

Kit Recovery of phosphopeptide standards Specificity*
1 2 3 Total
Sigma 59% 52% 74% 59% 50%
Competitor A 6% 19% 11% 13% 28%
Competitor B 39% 56% 17% 42% 28%
Competitor C 37% 65% 37% 46% 25%

Go now to PP0410 for your price and to order powerful phosphopeptide enrichment!
For additional information download the following posters:

Absolute Quantification (AQUA™) of Phosphorylation Levels in Biological Samples using Immobilized Metal Affinity Chromatography Enrichment (IMAC). (663 kb PDF)

A Novel Silica-Based Immobilized Metal Affinity Chromatography (IMAC) Technology for the Enrichment of Phosphopeptides.
(708 kb PDF)

Targeting the Phosphotyrosine Proteome Using an Immobilized Metal Affinity Chromatography Spin Column and MALDI Matrix Optimization for Mass Spectrometric Enhancement. (319 kb PDF)

Your library for understanding and applying the PhosphoProfile I technology

Title Author / Journal
Proteomic analysis of protein kinases by target class-selective pre-fractionation and tandem mass spectrometry. Wissing J, Jansch L, Nimtz M, Dieterich G, Hornberger R, Keri G, Wehland J, Daub H.
Mol Cell Proteomics 2006. Dec 27.
Phosphoproteomics toolbox: computational biology, protein chemistry and mass spectrometry. Hjerrild M, Gammeltoft S.,
FEBS Lett. 2006. Sep 4; 580(20): 4764-70. Epub 2006 Aug 4.
Switching the flip: protein phosphatase roles in signaling pathways. DeLong A.,
Curr Opin Plant Biol. 2006. Oct; 9(5): 470-7. Epub 2006 Aug 4. Review
Peptide-based phosphoproteomics with immobilized metal ion chromatography. Nuhse TS, Peck SC.,
Methods Mol Biol. 2006.; 323: 431-6.

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