Liquid chemiluminescent DNA pull-down assay to measure nuclear receptor-DNA binding in solution.
Electrophoretic mobility shift assays (EMSAs) are commonly used to investigate protein-DNA binding in vitro. However, EMSA can generate considerable amounts of undesirable waste, particularly when toxic compounds are examined. We therefore developed a novel in vitro protein-DNA binding assay called liquid chemiluminescent DNA pull-down assay, which is based on solution hybridization between digoxigenin-labeled DNA and glutathione S-transferase (GST)-fused DNA binding protein bound to glutathione-Sepharose beads.
Iwasaki
Toshiharu
T
Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan. tiwasaki@med.gunma-u.ac.jp
Miyazaki
Wataru
W
Rokutanda
Nana
N
Koibuchi
Noriyuki
N
D004247:DNA / Q000378:metabolism*; D008163:Luminescent Measurements / Q000379:methods*; D011485:Protein Binding; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D012996:Solutions
D018160:Receptors, Cytoplasmic and Nuclear; D012996:Solutions; D004247:DNA