Optimization of Peptide Density on Microarray Surface for Quantitative Phosphoproteomics

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  • SHIMOMURA Takashi
    Department of Applied Chemistry, Kyushu University
  • HAN Xiaoming
    Department of Applied Chemistry, Kyushu University
  • HATA Akito
    Department of Applied Chemistry, Kyushu University
  • NIIDOME Takuro
    Department of Applied Chemistry, Kyushu University Center for Future Chemistry, Kyushu University
  • MORI Takeshi
    Department of Applied Chemistry, Kyushu University Center for Future Chemistry, Kyushu University
  • KATAYAMA Yoshiki
    Department of Applied Chemistry, Kyushu University Center for Future Chemistry, Kyushu University

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Abstract

Procedures to determine the density of peptides immobilized on a glass surface for the quantitative detection of phosphorylated peptides for phosphoproteomic applications of peptide microarrays are described. Two kinds of representative fluorescent probe molecules, anti-phosphotyrosine antibody (AB) and phos-tag (PT), were examined to compare their ability for the quantitative detection of phosphorylated peptides. PT is a metal complex with a binding specificity to phosphorylated amino acids, and is much smaller in size than AB. Thus, PT is quantitatively bound to the phosphorylated peptides, even at a high immobilization density without steric hindrance, making them highly suited for future microarrays requiring smaller sized peptide spots for much higher throughput.

Journal

  • Analytical Sciences

    Analytical Sciences 27 (1), 13-17, 2011

    The Japan Society for Analytical Chemistry

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