Label-Free Electrophoretic Mobility Shift Assay (EMSA) for Measuring Dissociation Constants of Protein-RNA Complexes
نویسندگان
چکیده
منابع مشابه
Electrophoretic Mobility Shift Assay (EMSA).
Transcriptional regulation of gene expression is controlled through the binding of sequence-specific DNA-binding proteins (transcription factors) to the regulatory regions of genes. The exact gene expression program of a cell is determined by the spectrum of transcription factors present with the nucleus of a cell. The presence of these factors is dependent upon the cell type being examined and...
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The electrophoretic mobility shift assay (EMSA), or gel mobility shift assay, is a popular and powerful technique for the detection of RNA-protein interactions. It relies on the fact that naked RNA has certain mobility on nondenaturing gels, but if the RNA is bound by protein, the mobility of the RNA is reduced. Therefore, the binding of protein results in a characteristic upward shift of the R...
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The characterization of protein-nucleic acid interactions is essential not only for understanding the wide range of cellular processes they are involved in, but also the mechanisms underlying numerous diseases associated with the breakdown of regulatory systems. These include, but are far from being limited to, cell cycle disorders such as cancer and those caused by pathogenic agents that rely ...
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We describe a protocol for the fluorescent electrophoretic mobility shift assay improved for the quantitative analysis of protein-DNA complexes. Fluorescent-labeled oligonucleotide probes incubated with nuclear proteins were followed by electrophoresis. The signals for protein-DNA complexes were measured and normalized with fluorescent-labeled marker using fragment analysis software. This assay...
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The elongation phase of transcription by RNA polymerase II (RNAP II) is controlled by a carefully orchestrated series of interactions with both negative and positive factors. However, due to the limitations of current methods and techniques, not much is known about whether and how these proteins physically associate with the engaged polymerases. To gain insight into the detailed mechanisms invo...
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ژورنال
عنوان ژورنال: Current Protocols in Nucleic Acid Chemistry
سال: 2018
ISSN: 1934-9270
DOI: 10.1002/cpnc.70