Liquid chemiluminescent DNA pull-down assay to measure nuclear receptor-DNA binding in solution. (October 2008)

• Record Type: Journal Article
• Authors: Iwasaki, Toshiharu; Miyazaki, Wataru; Rokutanda, Nana; Koibuchi, Noriyuki
• Is Part Of: Biotechniques. Volume 45:Number 4(2008)
• Page Start: 445
• Page End: 448
• Publication Date: 2008-10
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/000112915
• Languages: English
• ISSNs: 0736-6205

Abstract

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.