Design of a fluorescent electrophoretic mobility shift assay improved for the quantitative and multiple analysis of protein-DNA complexes.
We describe a protocol for the fluorescent electrophoretic mobility shift assay improved for the quantitative analysis of protein-DNA complexes. Fluorescent-labeled oligonucleotide probes incubated with nuclear proteins were followed by electrophoresis. The signals for protein-DNA complexes were measured and normalized with fluorescent-labeled marker using fragment analysis software. This assay proved reliable measurement and multiple detection of DNA binding proteins.
Onizuka
Takuo
T
Biological Sciences Department, Toray Research Center, Inc., 1111 Tebiro, Kamakura, Kanagawa 248-8555, Japan. Takuo_Onizuka@trc.toray.co.jp
Endo
Sumiyo
S
Hirano
Masahiko
M
Kanai
Shozo
S
Akiyama
Hideo
H
D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods*; D005453:Fluorescence; D011485:Protein Binding; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D004247:DNA