[An infrared imaging system for detecting electrophoretic mobility shift of DNA-protein complexes].
OBJECTIVE
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.


METHODS
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.


RESULTS
One lane of DNA-protein complex was clearly presented, and the signal intensity increased along with the increment of the protein load.


CONCLUSIONS
This infrared imaging system can be used for EMSA for detecting the DNA-protein complex with high sensitivity efficiency and allows easy operation.
Wang
Yi-fei
YF
Department of Endocrinology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
Cai
De-hong
DH
Chen
Hong
H
Mo
Yong-yan
YY
Yi
Na
N
Xing
Fei-yue
FY
D001665:Binding Sites; D004247:DNA / Q000737:chemistry*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay / Q000295:instrumentation* / Q000379:methods; D005456:Fluorescent Dyes / Q000737:chemistry; D005786:Gene Expression Regulation; D006801:Humans; D007259:Infrared Rays; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs / Q000502:physiology; D011506:Proteins / Q000737:chemistry*
D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; D011506:Proteins; D004247:DNA