Abstract − Analytical Sciences, 27(1), 13 (2011).
Optimization of Peptide Density on Microarray Surface for Quantitative Phosphoproteomics
Takashi SHIMOMURA,* Xiaoming HAN,* Akito HATA,* Takuro NIIDOME,*,** Takeshi MORI,*,** and Yoshiki KATAYAMA*,**
*Department of Applied Chemistry, Kyushu University, 744 Moto-oka, Nishi, Fukuoka 819-0395, Japan
**Center for Future Chemistry, Kyushu University, 744 Moto-oka, Nishi, Fukuoka 819-0395, Japan
**Center for Future Chemistry, Kyushu University, 744 Moto-oka, Nishi, Fukuoka 819-0395, Japan
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.
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