Optimization of peptide density on microarray surface for quantitative phosphoproteomics

Takashi Shimomura, Xiaoming Han, Akito Hata, Takuro Niidome, Takeshi Mori, Yoshiki Katayama

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)

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. 2011

Original languageEnglish
Pages (from-to)13-17
Number of pages5
Journalanalytical sciences
Volume27
Issue number1
DOIs
Publication statusPublished - 2011

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

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