An infrared imaging system for detecting electrophoretic mobility shift of DNA-protein complexes.
- Author:
Yi-fei WANG
1
;
De-hong CAI
;
Hong CHEN
;
Yong-yan MO
;
Na YI
;
Fei-yue XING
Author Information
- Publication Type:Journal Article
- MeSH: Binding Sites; DNA; chemistry; DNA-Binding Proteins; chemistry; metabolism; Electrophoretic Mobility Shift Assay; instrumentation; methods; Fluorescent Dyes; chemistry; Gene Expression Regulation; Humans; Infrared Rays; Protein Binding; Protein Interaction Domains and Motifs; physiology; Proteins; chemistry
- From: Journal of Southern Medical University 2009;29(2):289-291
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSGILZ 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.
RESULTSOne lane of DNA-protein complex was clearly presented, and the signal intensity increased along with the increment of the protein load.
CONCLUSIONThis infrared imaging system can be used for EMSA for detecting the DNA-protein complex with high sensitivity efficiency and allows easy operation.
