Use of Modified Agarose Gel Electrophoresis to Resolve Protein-DNA Complexes for Electrophoretic Mobility Shift Assay
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...
متن کاملElectrophoretic mobility shift assays for RNA-protein complexes.
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...
متن کاملDesign of a fluorescent electrophoretic mobility shift assay improved for the quantitative and multiple analysis of protein-DNA complexes.
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 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 ...
متن کاملAgarose gel electrophoresis.
MATERIALS 6x Gel-loading buffer Agarose solutions (please see Step 3) DNA samples DNA size standards Samples of DNAs of known size are typically generated by restriction enzyme digestion of a plasmid or bacteriophage DNA of known sequence. Alternatively, they are produced by ligating a monomer DNA fragment of known size into a ladder of polymeric forms. DNA staining solution For a discussion of...
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ژورنال
عنوان ژورنال: BioTechniques
سال: 1998
ISSN: 0736-6205,1940-9818
DOI: 10.2144/98242bm09