Open AccessLiquid chemiluminescent DNA pull-down assay to measure nuclear receptor-DNA binding in solution
Author(s) -
Tadaaki Iwasaki,
Wataru Miyazaki,
Nana Rokutanda,
Noriyuki Koibuchi
Publication year - 2008
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000112915
Subject(s) - chemiluminescence , dna , microbiology and biotechnology , measure (data warehouse) , chemistry , biology , biochemistry , chromatography , computer science , database
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.