title: Characterization of protein-DNA interactions using protein microarrays.
abstract: Protein-DNA interactions (PDIs) are critical for many cellular processes. We present here a protocol for the identification of PDIs in vitro using protein microarray technology. The procedure involves double-stranding synthesized DNA oligonucleotides with a fluorescent-labeled primer, binding the labeled double-stranded DNA directly to the protein microarray, and analyzing binding of the resulting PDIs. This approach provides simultaneous identification of PDIs for thousands of proteins, and multiple carefully designed DNA probes can be tested in parallel, which enables a rapid mapping of PDIs on a proteome-wide scale.
authors.affiliation: Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
authors.forename: Shaohui
authors.initials: S
authors.lastname: Hu
authors.forename: Zhi
authors.initials: Z
authors.lastname: Xie
authors.forename: Seth
authors.initials: S
authors.lastname: Blackshaw
authors.forename: Jiang
authors.initials: J
authors.lastname: Qian
authors.forename: Heng
authors.initials: H
authors.lastname: Zhu
mesh_terms: D004247:DNA / Q000378:metabolism*; D005456:Fluorescent Dyes; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D011506:Proteins; D004247:DNA