Determination of nuclear transcription factor activity using a modified electrophoretic mobility shift assay

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...

Electrophoretic Mobility Shift Assay: Analyzing Protein – Nucleic Acid Interactions

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 ...

Analysis of factor interactions with RNA polymerase II elongation complexes using a new electrophoretic mobility shift assay

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...

An Optimized Protocol for Electrophoretic Mobility Shift Assay Using Infrared Fluorescent Dye-labeled Oligonucleotides.

Electrophoretic Mobility Shift Assays (EMSA) are an instrumental tool to characterize the interactions between proteins and their target DNA sequences. Radioactivity has been the predominant method of DNA labeling in EMSAs. However, recent advances in fluorescent dyes and scanning methods have prompted the use of fluorescent tagging of DNA as an alternative to radioactivity for the advantages o...

Determination of the Binding Site-Size of the Protein-DNA Complex by Use of the Electrophoretic Mobility Shift Assay