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Updated: Jun 5, 2026

Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer
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Optimization of peptide density on microarray surface for quantitative phosphoproteomics.

Takashi Shimomura1, Xiaoming Han, Akito Hata

  • 1Department of Applied Chemistry, Kyushu University, 744 Moto-oka, Nishi, Fukuoka 819-0395, Japan.

Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry
|January 15, 2011
PubMed
Summary
This summary is machine-generated.

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Phos-tag (PT) offers superior quantitative detection of phosphorylated peptides on microarrays compared to antibodies. This smaller molecule binds effectively at high densities, enabling higher throughput phosphoproteomic analysis.

Area of Science:

  • Biochemistry
  • Proteomics
  • Analytical Chemistry

Background:

  • Peptide microarrays are crucial for phosphoproteomic applications.
  • Accurate quantification of immobilized peptides is essential for reliable detection.
  • Current methods face challenges with high peptide densities and steric hindrance.

Purpose of the Study:

  • To describe procedures for determining peptide density on glass surfaces for quantitative detection.
  • To compare the efficacy of anti-phosphotyrosine antibody (AB) and phos-tag (PT) as fluorescent probes.
  • To evaluate their suitability for high-throughput phosphoproteomic analysis using peptide microarrays.

Main Methods:

  • Immobilization of peptides on a glass surface.
  • Quantitative detection using fluorescently labeled anti-phosphotyrosine antibody (AB).

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Last Updated: Jun 5, 2026

Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer
12:23

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Published on: August 2, 2018

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation
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An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation

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Quantitative Phosphoproteomics in Fatty Acid Stimulated Saccharomyces cerevisiae
15:41

Quantitative Phosphoproteomics in Fatty Acid Stimulated Saccharomyces cerevisiae

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  • Quantitative detection using fluorescently labeled phos-tag (PT), a metal complex.

    • Main Results:

    • Phos-tag (PT) demonstrated effective quantitative binding to phosphorylated peptides, even at high immobilization densities.
    • PT, being smaller than AB, avoids steric hindrance issues.
    • PT's binding characteristics are highly suitable for dense peptide microarrays.

    Conclusions:

    • Phos-tag (PT) is a superior probe for quantitative detection of phosphorylated peptides in high-density microarray applications.
    • PT enables higher throughput phosphoproteomic analysis due to its efficient binding.
    • This method advances the development of next-generation peptide microarrays.