Determination of nuclear transcription factor activity using a modified electrophoretic mobility shift assay
Yong Wang, Wenhua Huang, Daizhi Peng, Yuanbin Hu
Abstract
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Yong Wang, Wenhua Huang, Daizhi Peng, Yuanbin Hu
Abstract
Open-access reader
In this work, several major procedures of the electrophoretic mobility shift assay (EMSA) were modified including swift extraction of the nucleic protein, labeling of the probe and radioautography. The modified assay required shorter time, simplified the nucleic protein extraction, increased the radioactivity of the labeling probe, skipped the tedious process of gel drying, and produced clear images. Its results were comparable, reproducible and stable. It thus has merited for wide application.
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In this work, several major procedures of the electrophoretic mobility shift assay (EMSA) were modified including swift extraction of the nucleic protein, labeling of the probe and radioautography. The modified assay required shorter time, simplified the nucleic protein extraction, increased the radioactivity of the labeling probe, skipped the tedious process of gel drying, and produced clear images. Its results were comparable, reproducible and stable. It thus has merited for wide application.
Key concepts: Electrophoretic mobility shift assay, Nucleic acid, Electrophoresis, Chemistry, Extraction (chemistry), Chromatography, Molecular biology, Transcription factor