title: Optimization of peptide density on microarray surface for quantitative phosphoproteomics.
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.
authors.affiliation: Department of Applied Chemistry, Kyushu University, 744 Moto-oka, Nishi, Fukuoka 819-0395, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Shimomura
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Han
authors.forename: Akito
authors.initials: A
authors.lastname: Hata
authors.forename: Takuro
authors.initials: T
authors.lastname: Niidome
authors.forename: Takeshi
authors.initials: T
authors.lastname: Mori
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Katayama
mesh_terms: D005898:Glass / Q000737:chemistry; D010455:Peptides / Q000032:analysis* / Q000737:chemistry; D010748:Phosphopeptides / Q000032:analysis* / Q000737:chemistry; D040081:Protein Array Analysis / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D013499:Surface Properties
chemical_list: D010455:Peptides; D010748:Phosphopeptides