Electrophoretic Mobility Shift Assay Coupled with Immunoblotting for the Identification of DNA-Binding Proteins

DNA mobility shift assay coupled with SDS-PAGE for detection of DNA-binding proteins.

DNA mobility shift assay is widely performed to analyze sequence-specific DNA-binding proteins in cellular extracts (2), and the DNA-binding proteins can be identified by use of antibodies that induce further retardation of probes (supershift). Gel extracts from a shifted band contain not only DNA segments and DNA-binding proteins but also nonspecific proteins located at the same position in th...

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...

A proteomics approach for the identification of DNA binding activities observed in the electrophoretic mobility shift assay.

Transcription factors lie at the center of gene regulation, and their identification is crucial to the understanding of transcription and gene expression. Traditionally, the isolation and identification of transcription factors has been a long and laborious task. We present here a novel method for the identification of DNA-binding proteins seen in electrophoretic mobility shift assay (EMSA) usi...

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

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 ...