An infrared imaging system for detecting electrophoretic mobility shift of DNA-protein complexes / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To establish a new non-radioactive method for electrophoretic mobility shift assay (EMSA) to investigate the binding between glucocorticoid induced leucine zipper (GILZ) and peroxisome proliferator-activated receptor-gamma 2 (PPARgamma2) promoter oligonucleotides.</p><p><b>METHODS</b>GILZ protein prepared by prokaryotic expression was linked to PPARgamma2 promoter oligonucleotides end-labeled with IRDye 800 infrared dye. The DNA-protein complex was separated with non-denatured polyacrylamide gel and scanned with the Odyssey. Infrared Imaging System.</p><p><b>RESULTS</b>One lane of DNA-protein complex was clearly presented, and the signal intensity increased along with the increment of the protein load.</p><p><b>CONCLUSION</b>This infrared imaging system can be used for EMSA for detecting the DNA-protein complex with high sensitivity efficiency and allows easy operation.</p>